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J. Biol. Chem., Vol. 276, Issue 25, 22663-22674, June 22, 2001
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From the The gene encoding the murine
calcitonin receptor (mCTR) was isolated, and the exon/intron structure
was determined. Analysis of transcripts revealed novel cDNA
sequences, new alternative exon splicing in the 5'-untranslated
region, and three putative promoters (P1, P2, and P3). The
longest transcription unit is greater than 67 kilobase pairs, and the
location of introns within the coding region of the mCTR gene (exons
E3-E14) are identical to those of the porcine and human CTR genes. We
have identified novel cDNA sequences that form three new exons as
well as others that add 512 base pairs to the 5' side of the
previously published cDNA, thereby extending exon E1 to 682 base
pairs. Two of these novel exons are upstream of exon E2 and form a
tripartite exon E2 (E2a, E2b, and E2c) in which E2a is utilized by
promoter P2 with variable splicing of E2b. The third new exon (E3b')
lies between E3a and E3b and is utilized by promoter P3. Analysis of mCTR mRNAs has revealed that the three alternative promoters give rise to at least seven mCTR isoforms in the 5' region of the gene and
generate 5'-untranslated regions of very different lengths. Analysis by
reverse transcription-polymerase chain reaction shows that promoters P1
and P2 are utilized in osteoclasts, brain, and kidney, whereas promoter
P3 appears to be osteoclast-specific. Using transiently transfected
reporter constructs, promoter P2 has activity in both a murine kidney
cell line (MDCT209) and a chicken osteoclast-like cell line (HD-11EM),
whereas promoter P3 is active only in the osteoclast-like cell line.
These transfection data confirm the osteoclast specificity of promoter
P3 and provide the first evidence that the CTR gene is regulated in a
tissue-specific manner by alternative promoter utilization.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF333472 (mCTR E1/E2 genomic), AF333473 (mCTR
E3a/E3b' genomic), AF333474 (mCTR E3b genomic), AF333475 (mCTR E4
genomic), AF333476 (mCTR E5 genomic), AF333477 (mCTR E6 genomic), AF333478 (mCTR E7 genomic), AF333479 (mCTR E8a genomic), AF333480
(mCTR E8b genomic), AF333481 (mCTR E9 genomic), AF333482 (mCTR E10/E11
genomic), AF333483 (mCTR E12 genomic), AF333484 (mCTR E13 genomic),
AF333485 (mCTR E14 genomic).
Tissue-specific and Ubiquitous Promoters Direct the Expression of
Alternatively Spliced Transcripts from the Calcitonin Receptor
Gene*
§¶,
**,,,,, and§§
New England Baptist Bone and Joint
Institute, Beth Israel Deaconess Medical Center, Boston, Massachusetts
02115, § Harvard School of Dental Medicine, Boston,
Massachusetts 02115, and Harvard Medical School,
Boston, Massachusetts 02115
*
This work was supported by National Institutes of Health
Grants AR45421 (to D. L. G.) and DK46773 (to S. R. G.).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.
Present address: IDEC Pharmceuticals, Molecular Biology
Department, 11011 Torreyana Rd., San Diego, CA 92121.
§§
To whom correspondence should be addressed: Deborah L. Galson,
Beth Israel Deaconess Medical Center, New England Baptist Bone and
Joint Institute, Harvard Institutes of Medicine, Rm. 247, 4 Blackfan
Circle, Boston, MA 02115. Tel.: 617-667-0743; Fax: 617-975-5299;
E-mail: dgalson@caregroup.harvard.edu.
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