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J. Biol. Chem., Vol. 275, Issue 25, 18664-18669, June 23, 2000
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From the Institut National de la Santé et de la Recherche
Médicale, Unité 468, Hôpital Henri Mondor,
94010 Créteil Cedex, France
Received for publication, February 28, 2000, and in revised form, March 30, 2000
In humans, growth hormone receptor (GHR)
transcripts exist in two isoforms differing by the retention (GHRfl) or
exclusion (GHRd3) of exon 3, whereas in mice GHRfl is solely expressed. This species-specific expression pattern is believed to result from an
alternative splice event that, on the basis of conflicting data
obtained in humans, has been considered to be tissue-, developmentally, and/or individual-specific. To decipher the molecular basis of this
unusual trait, we isolated a 6.8-kilobase fragment spanning exon 3 from
individuals expressing GHRfl. Sequence analysis revealed the existence
of two 99% identical retroelements flanking this exon. Unexpectedly,
individuals expressing GHRd3 displayed a 2.7-kilobase deletion
involving exon 3, which most likely results from an ancestral homologous recombination between the two retroelements. The lineage of
these retroelements during primate evolution revealed the species specificity of the GHRd3 allele. These findings led us to propose a
model underlying the existence of the sole GHRfl allele in most species. Such a retrovirus-mediated alternative splice mimicry, which
clears up several as yet unexplained phenomena (i.e. the above-mentioned expression data, the Mendelian inheritance of GHR
expression patterns, and the deletion of nonconsecutive exons in
growth hormone resistant patients), represents a novel
physiological mechanism accounting for protein diversity between and
within species.
There is a variety of well documented mechanisms underlying the
complexity and the diversity of the structure and regulation of
eukaryotic nuclear genes during evolution. Several of these mechanisms
result from the presence of repetitive DNA sequences that form a
substantial fraction of these genomes (1), as well as from the
existence, in the great majority of genes coding for proteins, of
exonic and intronic sequences (2). Such a genomic organization accounts
for several kinds of DNA rearrangements (1, 3) and for mRNA
splicing (4). The latter corresponds to the precise excision of introns
from nascent transcripts followed by the ligation of exons. Alternative
splicing of primary transcripts represents a critical step in the
regulation of gene expression, generating different mature mRNAs
and thus different proteins from one gene (5). This mechanism can be
controlled in a cell type-specific manner, so that, depending on the
nature and/or the developmental stages of the tissues examined,
different mature mRNA species can be expressed; other alternative
splice events appear to be constitutive, with different mRNA
isoforms coexisting at constant ratios within the same cells (4).
The description of two different mature growth hormone receptor
(GHR)1 transcripts is a
particularly attractive example of an unusual phenomenon described as
the result of an alternative splice event that, in humans, has been
documented as being either constitutive or controlled in a cell
type-specific manner (6-12). The GHR protein is an integral cell
membrane molecule that, in humans, contains 638 residues (including a
membrane signal peptide of 18 residues); it consists of an
extracellular hormone-binding domain of 246 amino acids, a single
transmembrane domain, and a cytoplasmic domain of 350 residues (13).
Upon binding to the growth hormone, the GHR molecule can form
homodimers (14) that are essential to receptor activation (15, 16),
thereby mediating the well known biological effects of growth hormone.
The critical importance of the receptor in the control of body growth
was clearly demonstrated by the description of numerous GHR mutations
in patients with Laron syndrome, a severe growth hormone-resistant
short stature condition transmitted as an autosomal recessive trait
(17).
The human GHR gene is a single copy gene that spans 90 kb of the
5p13-p12 chromosomal region (6, 18). It contains nine coding exons
(numbered 2-10) and several untranslated exons: exon 2 codes for the
signal peptide, exons 3-7 encode the extracellular domain, exon 8 codes the transmembrane domain, and exons 9 and 10 code the cytoplasmic
domain (6). The existence of several GHR cDNA clones that diverge
within the coding region was first reported by Godowski et
al. (6). In one of these clones, the exon 3 sequence (consisting
of 66 nucleotides) was missing, in keeping with an alternative splice
event leading to either the retention or the exclusion of exon 3, corresponding to the full-length GHR isoform (GHRfl) or the exon
3-deleted isoform (GHRd3), respectively. Subsequent investigations have
yielded contradictory results. Both isoforms have been detected in
several human tissues, expressed either independently or
simultaneously. In the very first studies, a tissue-specific expression
pattern of GHRfl and GHRd3 was documented (7, 8), with GHRd3 mainly
expressed in the placental villi (7). However, other studies rather
indicated that the expression of these two GHR isoforms was specific
for each individual; different tissues obtained from the same
individual showed the same expression pattern (10, 11). This
individual-specific expression pattern of GHRfl and GHRd3 was found to
be maintained in long term cultures of fetal dermal fibroblasts (11).
Nevertheless, a cross-sectional study of a large number of human fetal
and postnatal tissues also suggested that the expression pattern of
GHRfl and GHRd3 may be developmentally regulated (11). Another study
showed that this alternative splice event resulted from an unusual
genetic polymorphism that is transmitted as a Mendelian trait and that
significantly alters splicing (12). Finally, the splicing of exon 3, whose functional consequences are poorly understood (8, 19), was also
suggested to be species-specific, because in mice the GHRfl transcript
is the only isoform detected (20).
To decipher the molecular basis of this unusual trait in humans, the
GHR genomic sequences surrounding exon 3 were studied in individuals
expressing GHRfl, GHRd3, or both isoforms. In addition, in an attempt
to provide an explanation for such species differences in the
expression of GHRd3 and GHRfl, similar studies were performed in
various species.
DNA and RNA Samples--
Human genomic DNA was isolated from
blood leukocytes by standard procedures. DNA samples were obtained from
150 unrelated control individuals. To assess GHRfl and GHRd3
expression, total RNA was extracted from Epstein-Barr virus-transformed
lymphocytes obtained in 11 of the 150 control individuals, using the
RNAplusTM protocol (Bioprobe Systems). The genomic DNA
samples of various primates including Lepilemur
(n = 2), Callimico (n = 8),
Cebus (n = 1), and Macaca mulatta
(n = 7) were kindly provided by Drs. Jacqueline
Levilliers (Institut Pasteur, Paris) and Florence Richard (Institut
Curie, Paris). African green monkey DNA was extracted from
the COS-7 cell line. The other monkey DNAs (gibbon, orangutan, gorilla,
and chimpanzee) were purchased from the European Collection of Cell Cultures.
Reverse Transcription PCR--
The GHR transcripts were
reverse-transcribed and subsequently amplified, as described (8).
Amplified products were analyzed by electrophoresis on a 6%
polyacrylamide gel stained with ethidium bromide.
Cloning of Exon 3-Flanking Sequences--
The genomic DNA from
an individual who expresses the full-length transcripts only was
digested with various restriction enzymes. Digested DNA fragments were
then self-ligated and subsequently used as templates in inverse PCR
(iPCR) assays. Briefly, 1 µg of genomic DNA was digested by one of
the following restriction enzymes: HindIII, MaeI,
BanII, or BglII. After ligation, iPCR was
performed in an Expand Long Template PCR System (Roche Molecular Biochemicals) with primers designed within exon 3, according to the
protocol recommended by the supplier. iPCR amplification products were
subsequently purified and directly sequenced by use of an ABI 373 automated DNA sequencer (Applied Biosystems). The location of the newly
identified sequence with respect to exon 3 was confirmed by direct
sequencing of PCR amplification products generated with primers
designed in this novel sequence. All sequences were determined on both strands.
Analysis of the GHR-Exon 3 Region in Humans and in Other Primate
Species--
PCR experiments were undertaken using various
combinations of primers to amplify the sequence surrounding exon 3 in
different DNAs obtained from individuals expressing GHRfl, GHRd3, or
both transcripts. The resulting PCR products were subsequently
sequenced. Finally, the existence of retroviral sequences (see below)
was investigated in the GHR-exon 3 region from the above-mentioned primate species, using the human GHR-specific primers
(GenBankTM accession numbers AF209078-AF209083 and
AF211184-211186).
Genotyping of GHR Exon 3--
To determine the genotype at the
GHR-exon 3 locus in a large population sample (i.e.
characterization of GHR DNA sequences associated with the expression of
GHRfl or GHRd3), a simple multiplex PCR assay was designed. This assay,
which was applied to the investigation of the 150 control genomic DNA
samples, was performed with primers G1, G2, and G3
(GenBankTM accession number AF155912), as follows: initial
step of denaturation of 5 min at 94 °C, followed by 35 cycles
consisting of 30 s at 94 °C, 30 s at 60 °C, and 1 min
30 s at 72 °C, followed by an extension period at 72 °C for
7 min. Amplification products were analyzed by electrophoresis on a 1%
agarose gel stained with ethidium bromide. The expected distribution of
the genotypes at the GHR-exon 3 locus was determined by means of the
Hardy and Weinberg law (21), which takes into account the allele
frequency of GHRfl and GHRd3 (see below) in our population sample.
Cloning of a 6.8-kb Fragment Surrounding Exon 3 of the Human GHR
Gene--
To test whether sequence differences in the exon 3-flanking
introns (i.e. introns 2 and 3) of the human GHR gene might
correlate with the exclusion or retention of exon 3 in GHR transcripts, we first amplified the exon 3-surrounding region of the genomic DNA
from an individual expressing the full-length GHR transcripts only by
means of iPCR. Indeed, the sequence of only 253 bp of the 3' end of
intron 2 and 71 bp of the 5' extremity of intron 3 has so far been
determined (12). These iPCR assays, performed on genomic DNA digested
by various restriction enzymes, led to the isolation of a 6.8-kb
fragment (Fig. 1A), which was
subsequently sequenced (GenBankTM accession number
AF155912). The identity of this DNA fragment as bona fide 5'
and 3' adjacent introns of exon 3 was confirmed in two ways. First, we
performed conventional PCR assays on the same DNA template with
different sets of primers whose sequences were derived from the newly
isolated sequence; as expected, the sequence of exon 3 was present in
all these amplified products. Second, restriction enzyme maps of the
PCR-amplified products surrounding exon 3 (Fig. 1A) were in
agreement with those obtained from the analysis of the products
generated through different iPCR assays.
The Human GHR Exon 3 Is Flanked by Two Retroviral Long Terminal
Repeat Fragments--
Complete sequence determination of this 6.8-kb
fragment revealed the existence of two 251-bp repeated elements (Fig.
1A). These repeats flank exon 3, with the 5' and the 3'
repeated elements located 577 bp upstream and 1821 bp downstream of the
exon, respectively. The nature of these elements was subsequently
determined by computer-assisted homology searches (RepeatMasker and
RepBase Web sites); they were found to be composed of a 171-bp-long
long terminal repeat (LTR) fragment from a human endogenous retrovirus
which belongs to the HERV-P family (22), followed by a 80-bp fragment
from a medium reiteration frequency MER4-type sequence (Fig.
1A) (23). The sequence of the two 251-bp-long copies
(referred to as 5' repeat and 3' repeat) are 99% identical, differing
in only three nucleotides at positions 14, 245, and 246 of the repeat.
More precisely, in all the alleles studied (n = 24),
the element located upstream from exon 3 carries a cytosine at position
14 and a thymine at positions 245 and 246, whereas the element located
downstream of the exon carries a guanine, a cytosine, and an adenine at
these positions. Extensive analysis of the entire 6.8-kb fragment by means of BLAST-N and RepeatMasker programs revealed the existence of
several other sequences of retroviral origin in the vicinity of the
exon; as shown in Fig. 1A, these sequences are members of
the MER family and of mammalian LTR transposons (23). In all the
alleles studied (n = 24), we found an identical genomic organization of the exon 3-surrounding region.
The LTR sequence located upstream from exon 3 derives from the class I
of the human endogenous retrovirus HERV-P (22, 24). This LTR element,
which is inserted in the opposite orientation to the transcription of
the GHR gene, displays the usual features of proviral inserted LTRs
(25); the sequence of both extremities, 5'-TGATATG..CATTTCA-3',
contains the universally conserved nucleotides common to all known
viral LTR (bold), as well as inverted repeated elements (underlined);
this retroelement also contains a potential polyadenylation signal
(AATAAA) and promoter sequence (TATAAA) (GenBankTM
accession number AF155912). In addition, the LTR element is flanked by
two short direct repeats differing at a single base (CCAT
(5'-TG..CA-3') CCAG), thereby indicating that this retroelement is
actually a solitary LTR. The 3' repeat contains a 171-bp-long sequence
identical to the 5' extremity of the 5' repeat. Computer-assisted translation of the total nucleotide sequence of the newly isolated 6.8-kb fragment did not reveal any sequence homology to gag,
pol, or env genes in the vicinity of these LTR elements.
A Homologous Recombination between Retroviral Repeated Elements
Results in the Genomic Deletion of Exon 3 in Individuals Expressing
GHRd3--
These findings led us to investigate genomic DNA samples
from individuals who express the GHRd3 isoform only. Somewhat
unexpectedly, exon 3 and its surrounding sequences could not be
amplified despite multiple attempts based on the use of different sets
and combinations of primers whose sequences were derived from the GHR
sequence located in between these two LTR-elements. However, using the same genomic DNAs as templates, a PCR amplification performed with a
sense primer (G1) located upstream of the 5' repeat and an antisense
primer (G2) located downstream of the 3' repeat yielded a 532-bp
fragment (Fig. 1B); this latter primer set amplified the
expected 3248-bp fragment when the genomic DNA from an individual who
expresses full-length GHR transcripts was used as a template (Fig.
1B). This result, which is consistent with a genomic
deletion in between primers G1 and G2, prompted us to sequence the
532-bp fragment. This analysis revealed the presence of a single
251-bp-long element and the absence of exon 3, thereby raising the
possibility that the deletion arose through recombination between the
two flanking retroelements (Fig. 1C). To address this issue,
the sequence of the DNA fragment surrounding the single LTR element was
determined (GenBankTM accession number AF210633) and
compared with that of a genomic DNA obtained from an individual who
expresses the GHRfl isoform only. This comparison showed perfect
matches between sequences located upstream from both the
GHRd3-associated retroelement and the 5' repeat found on GHRfl alleles
and between sequences located downstream of both the GHRd3-associated
retroelement and the 3' repeat found on GHRfl alleles. Restriction maps
of these sequences are depicted on Fig. 1C. The single
251-bp element that persists on the deleted allele was found to be
identical to the 3' copy identified on GHRfl alleles (i.e.
G, C, and A residues at positions 14, 245, and 246, respectively)
(n = 24). This arrangement of sequences is precisely
what would be expected if recombination between the 5' and 3' repeats
of the GHRfl allele occurred to delete a 2716-bp fragment, thereby
leaving behind a single solo LTR at the GHRd3 locus.
Distribution of GHRd3 and GHRfl in a Control Population
Sample--
To evaluate the allele frequencies of GHRd3 and GHRfl, we
developed a simple multiplex PCR assay based on the use of three primers: one antisense primer (G3) located in exon 3, and primer set
(G1 and G2), which brackets both the single LTR element of GHRd3
alleles and the two repeated elements of GHRfl (Fig.
2A). Under specific
experimental conditions, primers G1 and G2 allowed the amplification of
GHRd3 alleles only, whereas primers G1 and G3 amplified GHRfl alleles,
thereby allowing the accurate discrimination of the three possible
genotypes at this locus (i.e. homozygous GHRfl/GHRfl,
heterozygous GHRfl/GHRd3, and homozygous GHRd3/GHRd3) (Fig.
2B). A control population sample consisting of 150 unrelated individuals was investigated using this genotyping assay. The distribution of genotypes, which is presented in Table
I, follows the Hardy-Weinberg equilibrium
( Phylogenetic Analyses at the GHR-Exon 3 Locus--
To investigate
the phylogenesis of these retroelements on a time scale spanning
primate evolution, we performed PCR amplifications on various primate
genomic DNAs, using different sets of primers derived from the human
genomic sequence spanning exon 3. The absence of a retroelement
(HERV-P, MER-4, and/or mammalian LTR transposon sequences) from a
species was confirmed, in all cases, by PCR amplification of the
uninterrupted cellular target sequence. The retroviral sequences
identified in the vicinity of exon 3, which are compiled in Table
II, provide the following information.
First, the two retroelements that closely flank exon 3 (i.e.
MER49 and MaLR) are present in the hominoid lineage and in the genomes
of Old World monkeys and New World monkeys, whereas they are absent from the genome of Lepilemur, a prosimian primate. Second,
the HERV-P solitary LTR, which we identified in humans, is also present at the GHR-exon 3 locus of great apes and Old World monkeys but not at
that of New World monkeys and prosimians; this solitary LTR, which
shares common features with its human counterpart (i.e. inverted repeats, direct repeats, polyadenylation signal, and TATA
box), exhibits a sequence divergence with the human species of
5.8-6.4% and 2.0-3.9% for Old World monkeys and great apes, respectively. Third, the 251-bp-long 3' repeat identified in humans is
present in all the hominina subtribe (i.e. gorillas,
chimpanzees and humans), whereas it is missing altogether from all
other primate species, in which only an unoccupied integration site was
detectable.
We have described a new molecular mechanism to account for the
physiological expression, in humans, of two transcripts that differ by
the skipping of a coding exon. This phenomenon, which mimics
alternative splicing, results from a homologous recombination event
between retroviral sequences that flank the skipped exon. To our
knowledge, retroelement-induced effects of this kind have not been
reported before. As shown in this study focused on the analysis of the
GHR locus, the species specificity of these retroelements most likely
accounts for the documented species differences in the expression
pattern of GHR isoforms.
It is well established that in humans GHR transcripts are present in
two isoforms that differ by the retention or exclusion of exon 3, whereas in mice the GHRfl isoform is solely expressed. We have
demonstrated that in humans the GHRd3 isoform is actually transcribed
from a GHR allele that carries a 2.7-kb genomic deletion spanning exon
3, as compared with GHR alleles expressing GHRfl isoforms. More
precisely, we observed the association of exon 3 with two flanking
retroelements in the genomic DNA samples from individuals who solely
express GHRfl, whereas DNA samples from individuals who express GHRd3
contain only a single retroelement without exon 3. These observations
led us to assume that the 2.7-kb size difference documented between the
GHRd3 and GHRfl alleles is due to a deletion that occurred on an
ancestral GHRfl allele. As shown in the model depicted in Fig.
3, an intrachromosomal recombination
event most likely accounts for the generation of a GHRd3 allele from a
GHRfl template. Interchromosomal recombination between misaligned GHRfl
alleles was put aside as the basis for the exon 3 deletion; indeed,
such an event would also generate a reciprocal chromosome with an exon
3 duplication, this gene rearrangement having never been detected in
the population tested. Sequence analysis of 24 deleted alleles supports
the existence of a unique recombination breakpoint located within the
first 13 bases of the 251-bp repeated element (Fig. 3). In this model, the excision of exon 3 and its flanking sequences results from a
homologous recombination event between the two retroelements located on
the same GHRfl allele.
The identification of such an alternative splice mimicry clears up
several as yet unexplained phenomena. First, depending on the different
studies performed in humans since 1992 (7-12), the expression pattern
of GHRfl and GHRd3 has been considered to be tissue-, developmentally,
and/or individual-specific; in the light of the above-described
mechanism underlying GHRfl and GHRd3 expression, such conflicting
results are now easily reconcilable. Second, our results provide a
rational explanation for the Mendelian mode of inheritance of GHRfl and
GHRd3 expression patterns (12). Third, these findings also provide a
clear-cut explanation for the deletion of nonconsecutive exons
involving exons 3, 5, and 6 in patients with Laron syndrome (6); in
fact, the latter would rather be the product of a deletion of the
consecutive exons 5 and 6 on a GHRd3 allele. However, the amplification
of exon 3 in DNA samples from human tissues expressing the GHRd3
transcripts only (11, 12) appears not to be consistent with the
discovery of such a frequent genomic deletion of this exon found in 75 of the 300 GHR alleles of our population sample.
To determine the extent to which other species are able to generate
GHRd3 transcripts by means of a mechanism identical to that identified
in humans, we investigated the presence of these sequences at the
GHR-exon 3 locus from various primates. Because seven species from
hominoid to Old World monkeys carry the same solo LTR at the GHR locus
upstream from exon 3, we conclude that this LTR sequence derives from a
HERV-P proviral insertion that occurred in the germ line of a common
ancestor of the Old World lineages; in addition, because New World
monkeys and prosimii do not possess this solo LTR, we were able to
accurately date this insertion after the
Platyrrhini/Catarrhini split, thus about 25 million years
ago (26, 27). Strikingly, the retroelement located downstream of exon 3 in the human species (i.e. the 251-bp-long 3' repeat) is
present at the same locus only in gorillas and chimpanzees, thereby
demonstrating the recent origin of this integration event that occurred
after the divergence of Pongina and Hominina
(about 7 million years ago) (27). Several lines of evidence support the
hypothesis that the 3' repeat derives from the 5' repeat by means of a
duplication event. First, these highly homologous sequences share
identical 5' direct and inverted repeats. In addition, not only is the
3' repeat composed of a fragment of the HERV-P retroviral sequence, but
it also contains a perfect 40-bp-long copy of the MER4-type sequence
belonging to the 5' repeat that, by itself, is incapable of autonomous
retrotransposition. Furthermore, the low percentage of sequence change
between the 5' and 3' repeats within each species of the hominina
subtribe (1.2-2.0%) is consistent with the reference date of the
divergence of Pongina and Hominina (27).
Moreover, it is noteworthy that the 3' end of the repeated elements
contains a 8-bp-long sequence that is also present in the GHR gene of
all primate species studied so far (data not shown); it is therefore
tempting to speculate that this 8-bp-long cellular sequence, which is
located downstream of exon 3 at a position corresponding precisely to
the insertion site of the 3' repeat, may have favored the partial
duplication of the 5' repeat. Examination of GHRd3 alleles in the human
species led to the identification of a single retroelement that is
100% identical to the 3' repeat found on GHRfl alleles (Fig. 3); this
result, which indicates the absence of de novo mutation, is
consistent with a homologous recombination event that occurred very
recently during primate evolution. Taken together, these data are
consistent with the existence of the sole GHRfl allele in all primate
species except Hominina. Because the three species of the
Hominina subtribe carry both the 5' and 3' repeats, one
cannot rule out the possibility that in gorillas and chimpanzees, GHRd3
alleles could occur by means of a similar homologous recombination
event; this could be addressed by the analysis of DNA samples from a
large number of unrelated gorillas and chimpanzees.
A model that accounts for this sequence of events during molecular
evolution of the GHR locus is presented in Fig.
4. The proposed events include proviral
insertions leading to the MER49 and MaLR sequences prior to the HERV-P
proviral insertion upstream from exon 3, which occurred after the
emergence of New World monkeys. In this model, the latter event is
followed by (i) the generation of a HERV-P solo LTR through a
homologous recombination event between the 5' and 3' LTRs of the
proviral sequence, (ii) the partial duplication of the solo LTR within
the 3'-flanking region of exon 3, after the divergence of
Pongina and Hominina, and (iii) the homologous
recombination between the two repeated elements, which occurred
recently, at least in the human species, and led to the GHRd3 allele.
The GHR gene exists in species from distinct orders, such as
Primates, Carnivora, Cetartiodactyla,
Scandentia and Rodentia (28). In mice, the GHRfl
transcript is the only isoform detected (20), and remarkably, genomic
DNA does not hybridize to HERV-P sequences, as judged by Southern
blotting experiments (Ref. 24, and data not shown); this latter finding therefore further supports our molecular model inferring a key role of
these sequences of retroviral origin in the genesis of GHRd3
transcripts. Such a species-specific GHR structural difference, which
underlies the existence of the sole GHRfl allele in several species,
relates to the evolutionary differences of the mechanisms by which the
GHR gene gives rise to a soluble growth hormone-binding protein (29,
30), thereby making this gene a particularly interesting model to
investigate its encoded isoforms through evolution.
Species-specific Alternative Splice Mimicry at the Growth Hormone
Receptor Locus Revealed by the Lineage of Retroelements during Primate
Evolution
A NOVEL MECHANISM ACCOUNTING FOR PROTEIN DIVERSITY BETWEEN AND
WITHIN SPECIES*
§,,
ABSTRACT
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
EXPERIMENTAL PROCEDURES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
RESULTS
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
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Fig. 1.
Genomic organization of the human GHR locus
in the vicinity of exon 3. A, GHR exon 3 locus in an
individual expressing full-length GHR transcripts. Exon 3 is shown as a
black box. The LTR sequences originating from the human
endogenous retrovirus HERV-P are indicated by an arrow
(complete LTR sequence) or a rectangle (partial LTR
sequence). MER elements and the mammalian LTR transposon element are
indicated by ellipses and a square, respectively.
The 5' and 3' 251-bp repeated elements (indicated by 5'-R
and 3'-R, respectively) are illustrated by the gray
areas outlined by a rectangle (bottom). A
partial restriction enzyme map of the region is given. Bg,
BglII; B, BanII; H,
HindIII. B, electrophoresis of PCR-amplified
fragments spanning the GHR-exon 3 locus in individuals expressing
either GHRfl (fl) or GHRd3 (d3), and generated
with primers G1 and G2 under the following PCR conditions: denaturation
94 °C, 30 s; annealing 60 °C, 30 s; and elongation
72 °C, 4 min. The size marker (L) is the SmartLadder from
Eurogentec. C, schematic representation of human GHRfl and
GHRd3 alleles (drawn to scale), according to the data presented in Fig.
1B. A partial restriction enzyme map of the region is given.
Exon 3 is indicated by a black box. The repeated elements
are shown as gray boxes. Two repeated elements flank exon 3 on GHRfl alleles, whereas a single copy of the repeat is present on
GHRd3 alleles. Black arrowheads indicate the primers (G1 and
G2) used in the PCR experiments. The meeting point of the two dashed
lines shows the intragenic recombination breakpoint.
2 test = 2.5 < 3.84), with allele frequencies for GHRd3 and GHRfl of 25 and 75%, respectively. In a
smaller population (n = 11) in which both genomic DNA
and total RNA samples were available for study, this PCR assay showed a perfect correlation between the genotype (presence of GHRfl and/or GHRd3 alleles) and the expression pattern of one and/or two of these
isoforms (data not shown).
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Fig. 2.
Genotyping assay at the GHR-exon 3 locus. A, schematic representation of the human GHRfl
region including exon 3 (black box) and the repeated
elements (gray boxes). The GHRd3 allele contains a single
copy of the repeat (gray box). The position and orientation
of primers G1, G2, and G3 used in the multiplex PCR assay are indicated
by arrowheads. B, under specific experimental
conditions (i.e. denaturation 94 °C, 30 s; annealing
60 °C, 30 s; and elongation 72 °C, 1 min 30 s), primers
G1 and G2 allowed the amplification of GHRd3 alleles only, whereas
primers G1 and G3 amplify GHRfl alleles. The homozygous GHRfl,
heterozygous GHRfl/GHRd3 and homozygous GHRd3 genotypes are denoted by
fl, fl/d3, and d3, respectively.
Distribution of the genotypes at the human GHR-exon 3 locus in a large
population sample
Lineage of retroelements at the GHR-exon-3 locus during primate
evolution
DISCUSSION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
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Fig. 3.
Model for the origin of the genomic deletion
of exon 3. Exon 3 is shown as a black box. Gray
boxes denote the 5' and 3' repeats involved in the recombination
event. The two repeats are 99% identical, differing at only three
nucleotide positions, which were used to identify the location of the
breakpoint (×).
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Fig. 4.
Schematic representation of the proposed
phylogenetic model accounting for the structure of the GHR-exon 3 locus
in various primate species. Exon 3 is indicated by a black
box; the MER-type and MaLR proviral sequences are indicated by
empty circles and boxes, respectively. The HERV-P
provirus is presented in gray, with each arrow
denoting an LTR sequence.
The functional consequences of proviral integrations in mammalian
genomes were first reported as interfering widely with gene expression
when the cellular target sequence involves an exon (31), whereas it was
generally believed that insertions into noncoding regions would have
little or no effect on gene expression. This view, however, has been
challenged by the identification of mammalian sequences of retroviral
origin that are associated with a promoter activity (32-34) or that
induce a modification of mRNA splicing (35, 36). In contrast with
these situations, the present study reveals the existence of a
retrovirus-mediated mechanism underlying the physiological expression,
in humans, of two transcripts that differ by the skipping of a single
coding exon. This as yet unknown phenomenon, which mimics alternative splicing and accounts for protein diversity between and within species,
may represent one of the mechanisms underlying the expression of
structurally related isoforms at other loci.
| |
ACKNOWLEDGEMENTS |
|---|
We thank Drs. Jacqueline Levilliers and Florence Richard for supplying several monkey DNA samples.
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FOOTNOTES |
|---|
* This work was supported by an institutional grant from the INSERM.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF155912, AF210633, AF209078-209083, and AF211184-211186.
These authors contributed equally to this work.
§ Recipient of a fellowship from the Institut National de la Santé et de la Recherche Médicale.
¶ To whom correspondence should be addressed. E-mail: amselem@ im3.inserm.fr.
Published, JBC Papers in Press, April 7, 2000, DOI 10.1074/jbc.M001615200
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ABBREVIATIONS |
|---|
The abbreviations used are: GHR, growth hormone receptor; GHRfl, full-length GHR; GHRd3, GHR with exon 3 deleted; HERV, human endogenous retrovirus; LTR, long terminal repeat; MER, medium reiteration frequency sequence; kb, kilobase(s); PCR, polymerase chain reaction; iPCR, inverse PCR; bp, base pair(s).
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REFERENCES |
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TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
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|---|
| 1. | Charlesworth, B., Sniegowski, P., and Stephan, W. (1994) Nature 371, 215-220 |
| 2. | Berget, S. M. (1995) J. Biol. Chem. 270, 2411-2414 |
| 3. | Kazazian, H. H., Jr., and Moran, J. V. (1998) Nat. Genet. 19, 19-24 |
| 4. | Lopez, A. J. (1998) Annu. Rev. Genet. 32, 279-305 |
| 5. | Maniatis, T. (1991) Science 251, 33-34 |
| 6. | Godowski, P. J., Leung, D. W., Meacham, L. R., Galgani, J. P., Hellmiss, R., Keret, R., Rotwein, P. S., Parks, J. S., Laron, Z., and Wood, W. I. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 8083-8087 |
| 7. | Urbanek, M., MacLeod, J. N., Cooke, N. E., and Liebhaber, S. A. (1992) Mol. Endocrinol. 6, 279-287 |
| 8. | Sobrier, M. L., Duquesnoy, P., Duriez, B., Amselem, S., and Goossens, M. (1993) FEBS Lett. 319, 16-20 |
| 9. | Mercado, M., DaVila, N., McLeod, J. F., and Baumann, G. (1994) J. Clin. Endocrinol. Metab. 78, 731-735 |
| 10. | Wickelgren, R. B., Landin, K. L., Ohlsson, C., and Carlsson, L. M. (1995) J. Clin. Endocrinol. Metab. 80, 2154-2157 |
| 11. | Zogopoulos, G., Figueiredo, R., Jenab, A., Ali, Z., Lefebvre, Y., and Goodyer, C. G. (1996) J. Clin. Endocrinol. Metab. 81, 775-782 |
| 12. | Stallings-Mann, M. L., Ludwiczak, R. L., Klinger, K. W., and Rottman, F. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 12394-12399 |
| 13. | Leung, D. W., Spencer, S. A., Cachianes, G., Hammonds, R. G., Collins, C., Henzel, W. J., Barnard, R., Waters, M. J., and Wood, W. I. (1987) Nature 330, 537-543 |
| 14. | De Vos, A. M., Ultsch, M., and Kossiakoff, A. (1992) Science 255, 306-312 |
| 15. | Fuh, G., Cunningham, B., Fukunaga, R., Nagata, S., Goeddel, D., and Wells, J. (1992) Science 256, 1677-1680 |
| 16. | Duquesnoy, P., Sobrier, M. L., Duriez, B., Dastot, F., Buchanan, C. R., Savage, M. O., Preece, M. A., Craescu, C. T., Blouquit, Y., Goossens, M., and Amselem, S. (1994) EMBO J. 13, 1386-1395 |
| 17. | Amselem, S., Sobrier, M. L., Dastot, F., Duquesnoy, P., Duriez, B., and Goossens, M. (1996) Baillieres Clin. Endocrinol. Metab. 10, 353-369 |
| 18. | Barton, D. E., Foellmer, B. E., Wood, W. I., and Francke, U. (1989) Cytogenet Cell Genet. 50, 137-141 |
| 19. | Urbanek, M., Russell, J. E., Cooke, N. E., and Liebhaber, S. A. (1993) J. Biol. Chem. 268, 19025-19032 |
| 20. | Barnard, R., Southard, J. N., Edens, A., and Talamantes, F. (1993) Endocrinology 133, 1474-1477 |
| 21. | Connor, J., and Ferguson-Smith, M. (1993) Essential Medical Genetics , pp. 104-110, Blackwell Scientific Publications, Oxford |
| 22. | Boeke, J. D., and Stoye, J. P. (1997) in Retroviruses (Coffin, J. M. , Hughes, S. H. , and Varmus, H. E., eds) , pp. 343-435, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY |
| 23. | Smit, A. F. (1996) Curr. Opin. Genet. Dev. 6, 743-748 |
| 24. | Harada, F., Tsukada, N., and Kato, N. (1987) Nucleic Acids Res. 15, 9153-9162 |
| 25. | Chen, H. R., and Barker, W. C. (1984) Nucleic Acids Res. 12, 1767-1778 |
| 26. | Johnson, W. E., and Coffin, J. M. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 10254-10260 |
| 27. | Goodman, M. (1999) Am. J. Hum. Genet. 64, 31-39 |
| 28. | Rand-Weaver, M., Kaweauchi, H., and Ono, M. (1993) in Title (Schreibman, M. , Scanes, C. , and Pang, P., eds) , pp. 13-42, Academic Press, San Diego |
| 29. | Dastot, F., Sobrier, M. L., Duquesnoy, P., Duriez, B., Goossens, M., and Amselem, S. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 10723-10728 |
| 30. | Dastot, F., Duquesnoy, P., Sobrier, M. L., Goossens, M., and Amselem, S. (1998) Mol. Cell. Endocrinol. 137, 79-84 |
| 31. | Kazazian, H. H., Jr., Wong, C., Youssoufian, H., Scott, A. F., Phillips, D. G., and Antonarakis, S. E. (1988) Nature 332, 164-166 |
| 32. | Ting, C. N., Rosenberg, M. P., Snow, C. M., Samuelson, L. C., and Meisler, M. H. (1992) Genes Dev. 6, 1457-1465 |
| 33. | Schulte, A. M., Lai, S., Kurtz, A., Czubayko, F., Riegel, A. T., and Wellstein, A. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 14759-14764 |
| 34. | Bi, S., Gavrilova, O., Gong, D. W., Mason, M. M., and Reitman, M. (1997) J. Biol. Chem. 272, 30583-30588 |
| 35. | Seperack, P. K., Mercer, J. A., Strobel, M. C., Copeland, N. G., and Jenkins, N. A. (1995) EMBO J. 14, 2326-2332 |
| 36. | Cachon-Gonzalez, M. B., Fenner, S., Coffin, J. M., Moran, C., Best, S., and Stoye, J. P. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 7717-7721 |
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