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J Biol Chem, Vol. 274, Issue 27, 18847-18850, July 2, 1999
,From the Department of Molecular Physiology and Biophysics, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232-0615
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ABSTRACT |
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 TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
REFERENCES
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We demonstrate, utilizing a temperature
conditional mutant allele of the gene encoding TAF25p, that this
non-histone-like TBP-associated factor, which is shared between the
TFIID and SAGA complexes, is required for bulk mRNA gene
transcription by RNA polymerase II in vivo. Immunoblotting
experiments indicate that at the restrictive temperature, inactivation
of TAF25p function results in a reduction of the levels of numerous
TFIID and SAGA subunits, indicating its loss of function, like the
histone-like TAFs, causes degradation of the constituents of these two
multisubunit complexes. These data suggest that TAF25p plays a key
structural role in maintaining TFIID and SAGA complex integrity. This
is the first demonstration that a non-histone-like TAF is required for
continuous, high level RNA polymerase II-mediated mRNA gene transcription in living yeast cells.
There is ongoing debate about the exact role that
TAFIIs,1 a family
of conserved integral protein subunits of TFIID, play in transcription
by RNA polymerase II (see Refs. 1 and 2 for recent reviews). In
contrast to initial studies, recently published reports from a number
of laboratories indicate that at least a subset of TAFIIs,
the so-called histone-like TAFIIs (3), are absolutely
essential for ongoing high level total mRNA gene transcription in vivo (4-7). The histone-like TAFIIs examined
in these experiments are of particular interest, since it was also
demonstrated recently that these TAFIIs, TAF61p (histone
H2B-like), TAF60p (histone H4-like), and TAF17p (histone H3-like), are
present in both the TFIID and SAGA complexes (8).
The data from these studies has been used to formulate a model that
postulates that the histone-like TAFIIs form the structural "core" for both TFIID and SAGA complexes and when the integrity of
these particular TAFIIs is compromised, then the complexes dissociate and cease to function while the resulting free subunits are
rapidly degraded. At present it is not yet clear whether the RNAP II
transcription requirement for histone-like TAFII function derives from the association of these TAFIIs with the
TFIID, the SAGA, or both complexes. Circumstantial evidence implicates
the TFIID complex though, since all of the
non-TAFII-encoding genes that comprise the SAGA complex
(with the exception of TRA1 (9)) are non-essential genes,
while (with but one exception, TAF30) all of the TFIID
TAFIIs are encoded by essential genes. This question remains to be formally tested though.
Having previously cloned and characterized TAF25, we decided
to attempt to generate temperature-conditional alleles of the TAF25 gene, which we could use as a tool to examine the role
of TAF25p in RNAP II transcription. We felt it important to test whether or not the non-histone-like TAFII, TAF25p, which is
present in both TFIID and SAGA, is globally involved in total mRNA
gene transcription by RNA polymerase II in vivo. We
successfully generated temperature-conditional alleles of
TAF25 and one particular mutant allele caused yeast cells to
rapidly cease growth at the restrictive temperature. We characterized
this taf25ts mutant in detail with
regards to its effects both upon specific and global mRNA gene
transcription and TFIID/SAGA subunit protein integrity. Contrary to the
TFIID functional and structural organization models recently described
by others (reviewed in Ref. 2), we found that TAF25p function is
continuously required for high level mRNA gene transcription
in vivo. This is the first such demonstration that a
non-histone-like TAFII protein is continuously required for
mRNA gene transcription in vivo. We discuss how these
new data regarding TAF25p may be incorporated into current models of
TFIID structural organization and function.
Yeast and Bacterial Strains and Cell
Cultivation--
Escherichia coli strain XL-1 Blue (10) was
used for routine plasmid propagation. E. coli KC8
(CLONTECH) was used for amplification of
hydroxylamine mutagenized plasmid DNA. E. coli strain BL21 (11) was used for recombinant protein production (12). Yeast strain
YEK16 (12), genotype MATa leu2 ura3 trp1 suc2 ade2
lys2 taf25 Generation and Analysis of Temperature-sensitive Alleles of
TAF25--
Plasmid DNA was mutagenized following the general protocol
of Sikorski and Boeke (17). Details available upon request. Seven plasmids containing taf25ts alleles
screened true through this protocol. The entire TAF25 ORF
and ~200 bp of DNA both up- and downstream of the ORF were sequenced
for these seven mutants.
Preparation and Analysis of RNA--
Total yeast cell RNA was
prepared using a hot phenol extraction method as detailed previously
(18). RNAs were quantitated using UV absorbance and RiboGreen
fluorescence (Molecular Probes), and equivalent amounts of RNA (2 µg)
were analyzed by either slot blotting with a
32P-5'-end-labeled oligo(dT)20 probe (19) or by
standard RNA (Northern) blotting of agarose gel fractionated total RNA
(5 µg). RNA gel blots were probed using a PCR-amplified ORF of the
gene indicated in the legend to Fig. 3 that had been
32P-labeled by random priming. Hybridization and washing
steps were performed as described (20). S1 nuclease protection assays
were performed as described previously (15). Specific RNA signals were
detected by autoradiography and PhorphorImaging (Molecular Dynamics).
Preparation of Yeast WCE and Immunodetection of Specific
Proteins--
Yeast WCEs for immunological detection of TFIID and SAGA
constituents were prepared as described previously (12, 21, 22). Specific polyclonal antibodies recognizing TFIID and SAGA substituents were prepared in rabbits (Bethyl Laboratories, Montgomery, TX) using
1-2 mg of purified recombinant protein as immunogen for each rabbit.
IgGs were affinity-purified using antigen coupled to Sepharose CL-4B as
detailed previously (12, 21). Immunoblots were performed as described
previously (12, 21) using optimal, empirically determined dilutions of
specific polyclonal IgGs, typically 1/5000 to 1/10,000. In the case of
HA3-TAF25p, monoclonal antibody 12CA5 (Boerhinger Mannheim)
was used as well.
Hydroxylamine Efficiently Generated Temperature-sensitive Alleles
of TAF25--
By using hydroxylamine mutagenesis a number of distinct
taf25ts alleles were generated. We
obtained 7 out of ~20,000 yeast transformants that exhibited a
readily scorable temperature-conditional growth phenotype. None of
these mutants displayed a cold sensitive growth phenotype nor did they
exhibit a cdc-like phenotype after extensive incubation at the
non-permissive temperature.2
The locations of these mutations in TAF25 and the growth
properties of these seven yeast strains are shown in Fig.
1A. Yeast strains YEK25.34 and
YEK25.75 represent a missense mutation of codon glycine 101 to glutamic
acid (GGG to GAG), while strains YEK25.19, YEK25.26, YEK25.59, and
YEK25.92 were generated by missense mutation of glycine 101 to arginine
(GGG to AGG). Yeast strain YEK25.76 carries a nonsense mutation at
codon arginine 111 (CGA to UGA). All of these mutations are of the type
expected for hydroxylamine mutagenesis, which induces C to T and G to A
transition mutations (17). As shown in the lower part of
Fig. 1A, none of these strains grow well at 37 °C. Strain
YEK25.75 exhibited the greatest growth deficiency at 37 °C (Fig. 1
and data not shown) and was therefore selected for further
analysis.
All of these mutations map to a portion of TAF25p that is highly
conserved between disparate species (Schizosaccharomyces pombe, Arabidopsis thaliana, Drosophila
melanogaster, Caenorhabditis elegans, mouse,
human3). In fact, glycine 101 is absolutely conserved in seven of seven TAF25p orthologs, while
arginine 111 is conserved in six out of seven (in S. pombe
this residue is Lys rather than Arg). Additional mutagenesis studies
performed in our laboratory3 indicate that residues in this
highly conserved region of TAF25 are mutationally sensitive
and therefore, the high conservation of this amino acid sequence is
functionally relevant. The region of homology among these TAF25p
proteins extends from upstream of glycine 101 essentially to the C
terminus of the molecule and sequences downstream of arginine 111 are
essential for yeast cell viability.3 By rights then, if all
these sequences are essential for TAF25p function, then YEK25.76 would
be expected to be non-viable. We hypothesized that this strain can
grow, at least at 22 °C, because the nonsense mutation at
TAF25 ORF codon 111 is suppressed at a frequency sufficient
to provide levels of TAF25p adequate for growth. To test this
hypothesis we generated a yeast strain, which carried a version of
TAF25 in which three consecutive stop codons were inserted
at codons 111, 112, and 113. As would be expected if our suppression
model is correct, this strain was inviable when the triply prematurely
terminated TAF25 gene was the sole source of TAF25p in the
cell.3
mRNA Synthesis Rapidly Ceases in Cells Carrying a
taf25ts Allele at the Non-permissive
Temperature--
When cultures of YEK25.75 cells, which carry a
taf25ts allele, are shifted from 22 to
37 °C, their growth rate rapidly decreases and within 6 h
ceases altogether (Fig. 1B). We examined RNA synthesis in
TAF25 and taf25ts cells grown
at 22 and 37 °C for various lengths of time in order to assess the
effects of this temperature shift upon RNA synthesis. As controls,
separate, independent cultures of cells carrying rpb1-1
(which encodes the largest subunit of RNA polymerase II), tsm1-1 (which encodes the TFIID subunit TAF150p), or
tbpts-1 (which encodes TBP) mutant
genes were also utilized and processed for RNA in parallel. Polymerase
II-mediated mRNA gene transcription has been shown to be quite
sensitive to loss of both Rpb1p (13) and TBP (15) function in
vivo, while RNA polymerase II transcription in vivo
appears insensitive to the loss of TAF150p (23).
The results of our analyses examining total poly(A)+
mRNA are presented in Fig.
2A. It is clear from these
data that total mRNA synthesis in cells bearing the
taf25ts mutation decreases almost as
rapidly after a temperature shift as does mRNA biosynthesis in the
yeast strain carrying the rpb1-1 mutation, a benchmark
strain for such analyses. Total mRNA synthesis in
tbpts-1 cells is, as expected, also
sensitive to a temperature shift while mRNA synthesis is
insensitive to temperature in cells bearing either the wild type
TAF25 gene or the tsm1-1 mutation. Importantly the decrease in total mRNA synthesis in the
taf25ts cells is nearly identical to the
rate of loss of mRNA biosynthesis in the
tbpts-1 cells, indicating as direct
of an involvement of TAF25p in RNA polymerase II-mediated transcription
as that played by TBP. This loss of total mRNA synthesis in the
taf25ts cells is not due to general cell
death as >95% of the cells remain viable after 4 h at 37 °C
(data not shown).
The decrement in RNA synthesis in taf25ts
cells is restricted to mRNA biosynthesis as tRNA synthesis, which
is catalyzed by RNA polymerase III, was unaffected as measured by S1
nuclease protection assays (Fig. 2B). As expected, tRNA
synthesis decreases rapidly in
tbpts-1 cells (15). The rapid drop in
total mRNA gene transcription observed in the slot blot analyses of
poly(A)+ mRNA is also observed if the levels of
specific mRNAs is examined by RNA (Northern) blotting (Fig.
2C). mRNA levels decrease specifically in
taf25ts cells shifted to the
non-permissive temperature at rates roughly proportional to the
half-lives of the individual mRNAs with six of the seven genes
analyzed (compare the loss of mRNAs in rpb1-1 and
taf25ts cells, Fig. 2C). Only
the expression of MET19 does not decrease at an equivalent
rate in taf25ts versus
rpb1-1 cells. However, mRNAMET19 levels are
reduced at later points (see 2 and 3 h, Fig. 2C), most
likely an indirect effect due to loss of TBP or other TAFII protein levels (see below). In total, these data clearly demonstrate that continued TAF25p function is acutely required for ongoing high
level RNA polymerase II-mediated mRNA gene transcription in living
yeast cells.
TFIID and SAGA Subunit Levels Are Differentially Degraded in
taf25ts Cells after a Shift to Non-permissive
Temperature--
As detailed in the Introduction, a model emanating
from the recent studies of TAFII function in TFIID argues
that the reason that the integrity of the histone-like
TAFIIs, TAF61p, TAF60p, and TAF17p, is so crucial for TFIID
activity is because these three TAFIIs form the
"structural core" of the complex. Once the integrity of this TFIID
core is compromised the complex dissociates and the free, non-complexed
subunits are degraded. In order to test whether a comparable situation
is operative in the case of the non-histone-like TAF25p, we examined
the levels of various TFIID and SAGA constituents by immunoblotting
SDS-PAGE fractionated WCEs prepared from both
taf25ts and TAF25 cells as a
function of time after a shift to 37 °C. These analyses, shown in
Fig. 3, were performed with aliquots of
the same cells used for the RNA experiments of Fig. 2 so that the
results are directly comparable. Although such chemiluminescence experiments are difficult to quantitate with great precision, it is
clear from these immunoblots that all of the various proteins analyzed
do specifically decrease with time in the
taf25ts cells. However, the rate of loss
of the different proteins is variable, falling roughly into three
classes. We have sorted these changes relative to the drop off in
mRNA synthesis in the taf25ts cells
where we observed a decrease of
To incorporate these new data regarding the non-histone-like TAF,
TAF25p, into current models of TFIID organization (see Introduction), we propose that, in addition to interactions between histone-like TAFs,
TAF25p-TAF25p interactions are critical for maintaining TFIID
structure, organization, and ultimately TFIID function. For example,
when TAF25p-TAF25p interactions are disrupted in YEK25.75 cells, which
carry the taf25ts mutant allele, overall
TFIID structure is disrupted and free TAFII subunits are
then degraded if they are not stably incorporated into TFIID. This
proposal is based upon our current data regarding the deleterious
effects of TAF25p inactivation on RNA polymerase II transcription and
TFIID subunit stability, our previously published data (12), which
indicates that TAF25p can interact with high affinity with itself both
in vitro and in vivo, and data that indicate
TAF25p is present in superstoichiometric amounts in a highly purified
TFIID fraction.2 This model can also be applied to SAGA
structure, organization, and function. Our data does not allow us to
definitively distinguish between loss of TFIID or SAGA function as the
contributing factor to the loss of bulk mRNA synthesis in
taf25ts cells. For reasons discussed
above, however, it is likely that loss of TFIID function is the major
contributing factor. Studies are currently in progress in our
laboratory to directly investigate these and other aspects of TFIID
structure and function using a combination of additional biochemical
and genetic analyses. Such studies will prove crucial to dissecting the
complex process of TFIID-mediated RNA polymerase II gene transcription.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
REFERENCES
MATERIALS AND METHODS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
REFERENCES
1::TRP1 pRS416-TAF25WT, was used for
transformation and plasmid shuffle testing of the ability of
mutagenized TAF25 alleles to grow at permissive and
non-permissive temperatures (see details below). Yeast strains carrying
rpb1-1, tsm1-1, and tbpts-1 mutant alleles have been
described previously (13-15). Yeast were grown in appropriate rich
(YPAD) or selective (SC) medium as required (16).
RESULTS AND DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
REFERENCES
View larger version (26K):
[in a new window]
Fig. 1.
Hydroxylamine generates mutations in
TAF25 that induce a temperature-sensitive growth
phenotype. A, hydroxylamine was used to generate random
mutations in TAF25. The NH2OH-mutagenized
TAF25 gene on a HIS3-marked CEN/ARS
plasmid (pRS413) was introduced into a yeast strain carrying a
taf25 null deletion allele in the chromosome and a
URA3-marked TAF25 gene on a CEN/ARS
plasmid (pRS416). The chromosomal null mutant was uncovered by plasmid
shuffle by plating on 5-FOA as detailed under "Material and
Methods." The ability of the mutagenized TAF25 gene to
support growth was monitored by plating on YPAD and incubation at
permissive (22 °C) and non-permissive (37 °C) temperatures.
Growth of various taf25ts mutant strains
is indicated in the photograph of the resulting plates. The codon
locations and the mutations in TAF25 that induce
temperature-sensitive growth are shown. DNA sequencing of the complete
TAF25p ORF indicated that these were the only mutations introduced into
TAF25. Strain YEK25.75, marked with an asterisk,
exhibited the most severe growth defect at 37 °C. B,
yeast strains carrying either a WT TAF25 allele or the
taf25ts allele were grown in YPAD liquid
culture at the permissive temperature (22 °C). One-half of the
culture was shifted to the non-permissive temperature (37 °C) at the
point indicated by the arrow. Growth was monitored by
measuring absorbance (light scattering) at 600 nm. These strains
exhibited equivalent growth properties whether growth was monitored by
A600 or cell counting (not shown). As controls,
cultures of yeast strains separately carrying rpb1-1,
tbpts-1, and tsm1-1
temperature-sensitive mutant genes were grown and manipulated similarly
(not shown). 
, TAF25 at 22 °C; 
, TAF25
at 37 °C; 
, taf25ts at 22 °C;
, taf25ts at 37 °C.
View larger version (47K):
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Fig. 2.
Characterization of RNA synthesis in the
taf25ts mutant. A,
total RNA was extracted from cells carrying TAF25,
taf25ts, rpb1-1,
tbpts-1, and tsm1-1
mutations as indicated, and the RNAs were processed to measure total
poly(A)+ mRNA content via slot blotting as detailed
under "Materials and Methods." Total poly(A)+ mRNA
levels were detected both by autoradiography and imaging; the
autoradiogram is shown. Labels indicate both the times at which the
cultures were sampled and the relevant genotype. The corresponding
percent control values at different times are: rpb1-1, 100, 9.4, 5.5, 4.2, 3.4, 3.2%; taf25ts, 100, 40.7, 45.4, 28.1, 14.2, 15.2%;
tbpts-1, 100, 27.8, 22.1, 23.3, 16.7, and 12.6% at t = 0, 0.25, 0.5, 1, 2, and 3 h
post-temperature shift, respectively. Experiments were performed in
triplicate with S.E. between ±1 and 10%. Similar results were
obtained in a second independent set of experiments. B, the
total RNA samples of A were reanalyzed for tRNA biosynthesis
by S1 nuclease protection as detailed under "Materials and
Methods"; labels are the same as in A. The corresponding
percent control values at different times are: TAF25, 100, 83, 101%; taf25ts, 100, 91, 73%;
tbpts-1, 100, 64, 43% at
t = 0, 1, and 3 h post-temperature shift,
respectively. C, the same RNA preparations analyzed in
A and B were separated by denaturing agarose gel
electrophoresis and probed for specific mRNA content as detailed
under "Materials and Methods." The specific mRNA species
analyzed (ACT1, DED1, ENO2, MET19, PCL1, RAD23, and
RPS5) are indicated by the labels. The corresponding percent
control values at different times are: for ACT1 mRNA in
the TAF25 strain, 100, 89, 105, 78%; in the
taf25ts strain, 100, 40.1, 8.9, 2.3%; in
the rpb1-1 strain, 100, 23.6, 3.7, 0.1%; for
ENO2 mRNA in the TAF25 strain, 100, 68.9, 73.6, 74.7%; in the taf25ts strain, 100, 58.1, 31.3, 11.8%; in the rpb1-1 strain, 100, 75, 30.9, 8.1% at t = 0, 1, 2, and 3 h post-temperature
shift, respectively. Similar results were obtained in a second
independent set of experiments.
70-80% in the first hour after the
temperature shift. This is obviously the time interval most relevant,
at least for initial discussion. TAFIIs 150p, 130p, 90p,
61p, 60p, 47p, 40p, 19p, and 17p all decrease by about 50% in the
first hour after the temperature shift. TAFIIs 67p and 25p
levels decrease more slowly, showing significant decreases only after
2 h at 37 °C. TAF25p, expressed from an HA3-tagged gene (see "Materials and Methods"), was analyzed using both
polyclonal anti-TAF25p IgG and the 12CA5 anti-HA mAb with comparable
results (data not shown). TBP (a TFIID subunit), Gcn5p (the acetylase subunit of the SAGA complex), and Ada2p (a SAGA subunit, not shown) behave similarly to TAF25p and TAF67p (Fig. 3). The levels of TAF30p,
an apparent subunit of multiple distinct complexes in addition to TFIID
(24, 25), do not significantly decrease over the time course
analyzed. Clearly, inactivation of TAF25p via temperature shift results
in the inactivation, through protein degradation, of the multiple
constituents of both TFIID and SAGA complexes. This phenomenon
has been seen by others in comparable analyses (4-7), although, as
stated in the Introduction, this inactivation-directed protein
degradation has been ascribed to abrogation of crucial structural
interactions between the histone-like TAFIIs.
View larger version (70K):
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Fig. 3.
Characterization of TFIID and SAGA protein
subunit levels in TAF25 and
taf25ts cells at various times
after a shift to non-permissive growth conditions. Cells taken
from the cultures analyzed in the experiments of Fig. 2 were harvested
and total WCE proteins prepared as detailed under "Materials and
Methods." These protein samples were fractionated by SDS-PAGE,
blotted to polyvinylidene difluoride membranes, and specific proteins,
as indicated, were detected by immunostaining with affinity-purified
polyclonal IgGs and chemiluminescence as detailed under "Materials
and Methods." In the case of certain TAFII proteins (see
TAF47p for example), protein levels were reduced at 0 h in
taf25ts cells as compared with the 0 h point from TAF25 cells.
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ACKNOWLEDGEMENTS |
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We thank our laboratory colleagues for their constructive criticism and advice throughout the course of these studies as well as for their generous sharing of reagents and strains. We thank K. Struhl, J. Haber, and R. Young for gifts of strains and plasmids. We also thank S. Berger and D. Allis for providing Ada2p and Gcn5p E. coli expression plasmids.
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FOOTNOTES |
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* This work was supported in part by National Institutes of Health Grant GM52461.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.
Received partial support from National Institutes of Health
Training Grant DK07563-09.
§ To whom correspondence should be addressed. Fax: 615-322-7236; E-mail: tony.weil{at}mcmail.vanderbilt.edu.
2 S. Sanders, unpublished results.
3 E. Klebanow, unpublished results.
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ABBREVIATIONS |
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The abbreviations used are: TAFs, TBP-associated factors; TAFII, RNA polymerase II-specific TAF; ts, temperature-sensitive; WT, wild type gene; GTF, general transcription factor; PIC, preinitiation complex; RNAP II, nuclear DNA-dependent RNA polymerase II; 5-FOA, 5-fluoroorotic acid; YPAD, yeast extract peptone dextrose medium supplemented with adenine; SC, synthetic complete medium; WCE, whole cell extract; ORF, open reading frame; SAGA, Spt, Ada, Gcn5 acetylase complex; PCR, polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis.
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REFERENCES |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
REFERENCES
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L. S. Martel, H. J. Brown, and A. J. Berk Evidence that TAF-TATA Box-Binding Protein Interactions Are Required for Activated Transcription in Mammalian Cells Mol. Cell. Biol., April 15, 2002; 22(8): 2788 - 2798. [Abstract] [Full Text] [PDF]
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R. J. Durso, A. K. Fisher, T. J. Albright-Frey, and J. C. Reese Analysis of TAF90 Mutants Displaying Allele-Specific and Broad Defects in Transcription Mol. Cell. Biol., November 1, 2001; 21(21): 7331 - 7344. [Abstract] [Full Text] [PDF]
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J. Kirchner, S. L. Sanders, E. Klebanow, and P. A. Weil Molecular Genetic Dissection of TAF25, an Essential Yeast Gene Encoding a Subunit Shared by TFIID and SAGA Multiprotein Transcription Factors Mol. Cell. Biol., October 1, 2001; 21(19): 6668 - 6680. [Abstract] [Full Text] [PDF]
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S. M. Kraemer, R. T. Ranallo, R. C. Ogg, and L. A. Stargell TFIIA Interacts with TFIID via Association with TATA-Binding Protein and TAF40 Mol. Cell. Biol., March 1, 2001; 21(5): 1737 - 1746. [Abstract] [Full Text]
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Y.-G. Gangloff, S. L. Sanders, C. Romier, D. Kirschner, P. A. Weil, L. Tora, and I. Davidson Histone Folds Mediate Selective Heterodimerization of Yeast TAFII25 with TFIID Components yTAFII47 and yTAFII65 and with SAGA Component ySPT7 Mol. Cell. Biol., March 1, 2001; 21(5): 1841 - 1853. [Abstract] [Full Text]
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Z. Chen and J. L. Manley Robust mRNA Transcription in Chicken DT40 Cells Depleted of TAFII31 Suggests Both Functional Degeneracy and Evolutionary Divergence Mol. Cell. Biol., July 15, 2000; 20(14): 5064 - 5076. [Abstract] [Full Text]
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O. Wolstein, A. Silkov, M. Revach, and R. Dikstein Specific Interaction of TAFII105 with OCA-B Is Involved in Activation of Octamer-dependent Transcription J. Biol. Chem., May 26, 2000; 275(22): 16459 - 16465. [Abstract] [Full Text] [PDF]
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 L. Kuras, P. Kosa, M. Mencia, and K. Struhl TAF-Containing and TAF-Independent Forms of Transcriptionally Active TBP in Vivo Science, May 19, 2000; 288(5469): 1244 - 1248. [Abstract] [Full Text]
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S. L. Sanders and P. A. Weil Identification of Two Novel TAF Subunits of the Yeast Saccharomyces cerevisiae TFIID Complex J. Biol. Chem., April 28, 2000; 275(18): 13895 - 13900. [Abstract] [Full Text] [PDF]
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Y. Tsukihashi, T. Miyake, M. Kawaichi, and T. Kokubo Impaired Core Promoter Recognition Caused by Novel Yeast TAF145 Mutations Can Be Restored by Creating a Canonical TATA Element within the Promoter Region of the TUB2 Gene Mol. Cell. Biol., April 1, 2000; 20(7): 2385 - 2399. [Abstract] [Full Text]
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G. Mengus, Y.-G. Gangloff, L. Carre, A.-C. Lavigne, and I. Davidson The Human Transcription Factor IID Subunit Human TATA-binding Protein-associated Factor 28 Interacts in a Ligand-reversible Manner with the Vitamin D3 and Thyroid Hormone Receptors J. Biol. Chem., March 31, 2000; 275(14): 10064 - 10071. [Abstract] [Full Text] [PDF]
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S. Georgieva, D. B. Kirschner, T. Jagla, E. Nabirochkina, S. Hanke, H. Schenkel, C. de Lorenzo, P. Sinha, K. Jagla, B. Mechler, et al. Two Novel Drosophila TAFIIs Have Homology with Human TAFII30 and Are Differentially Regulated during Development Mol. Cell. Biol., March 1, 2000; 20(5): 1639 - 1648. [Abstract] [Full Text]
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Y.-G. Gangloff, S. Werten, C. Romier, L. Carré, O. Poch, D. Moras, and I. Davidson The Human TFIID Components TAFII135 and TAFII20 and the Yeast SAGA Components ADA1 and TAFII68 Heterodimerize to Form Histone-Like Pairs Mol. Cell. Biol., January 1, 2000; 20(1): 340 - 351. [Abstract] [Full Text]
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 P. B. Komarnitsky, B. Michel, and S. Buratowski TFIID-specific yeast TAF40 is essential for the majority of RNA polymerase II-mediated transcription in vivo Genes & Dev., October 1, 1999; 13(19): 2484 - 2489. [Abstract] [Full Text]
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J. C. Reese, Z. Zhang, and H. Kurpad Identification of a Yeast Transcription Factor IID Subunit, TSG2/TAF48 J. Biol. Chem., June 2, 2000; 275(23): 17391 - 17398. [Abstract] [Full Text] [PDF]
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M. Lemaire and M. A. Collart The TATA-binding Protein-associated Factor yTafII19p Functionally Interacts with Components of the Global Transcriptional Regulator Ccr4-Not Complex and Physically Interacts with the Not5 Subunit J. Biol. Chem., August 25, 2000; 275(35): 26925 - 26934. [Abstract] [Full Text] [PDF]
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J. Wang and G. Dreyfuss A Cell System with Targeted Disruption of the SMN Gene. FUNCTIONAL CONSERVATION OF THE SMN PROTEIN AND DEPENDENCE OF Gemin2 ON SMN J. Biol. Chem., March 23, 2001; 276(13): 9599 - 9605. [Abstract] [Full Text] [PDF]
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H. Mitsuzawa, H. Seino, F. Yamao, and A. Ishihama Two WD Repeat-containing TATA-binding Protein-associated Factors in Fission Yeast That Suppress Defects in the Anaphase-promoting Complex J. Biol. Chem., May 11, 2001; 276(20): 17117 - 17124. [Abstract] [Full Text] [PDF]
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A. Rashevsky-Finkel, A. Silkov, and R. Dikstein A Composite Nuclear Export Signal in the TBP-associated Factor TAFII105 J. Biol. Chem., November 21, 2001; 276(48): 44963 - 44969. [Abstract] [Full Text] [PDF]
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