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Article: Intron-exon structure, alternative use of promoter and expression of the mouse collagen X gene, Col10a-1

TitleIntron-exon structure, alternative use of promoter and expression of the mouse collagen X gene, Col10a-1
Authors
Issue Date1993
PublisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/EJB
Citation
European Journal Of Biochemistry, 1993, v. 213 n. 1, p. 99-111 How to Cite?
AbstractThe entire mouse collagen X gene (Col10a-1) has been isolated. The gene is composed of three exons and two introns spanning 7.0 kb of the DNA sequence. Exons 2 and 3 together encode 15-bp of 5' untranslated sequence, a 2040-bp open reading frame and an 895-nucleotide 3' noncoding region. In the 5' flanking region of the gene, two consensus TATA-box sequences were found. Identification of the first exon by ribonuclease-protection assays and the determination of the 5' end of Col10a-1 mRNA transcripts by primer-extension analyses show that the more 3' TATA box is probably predominantly used and that there are at least three transcription start sites in the exon 1 sequence 3' to this, resulting in 5' untranslated regions of 78, 77 and 55 nucleotides. By means of rapid amplification of cDNA ends by polymerase chain reaction, an additional mRNA species was detected which overlapped the other Col10a-1 transcripts, including the 3' TATA box sequence, giving a 5' untranslated sequence of approximately 235 bases. This layer transcript starts approximately 20 bp 3' to the more 5' TATA box. The data suggest alternative use of promoters and transcription starts for the Col10a-1 gene. Comparison of the combined nucleotide and deduced amino acid sequences of exons 2 and 3 with chicken, bovine and human collagen X genes, showed a high degree of similarity indicating conservation of this gene throughout evolution. Mouse Col10a-1 mRNA was shown to be approximately 3.0 kb and the pepsinized protein. as detected by SDS/PAGE, was ~45 kDa. The mRNA and protein sizes correlate with that predicted by the open reading frame. Reverse-transcription polymerase chain reaction assays indicate that the mouse collagen X gene is first expressed at 13.5 days post coitum, temporally preceding the onset of endochondral ossification. In agreement with the generally accepted association of type-X collagen with endochondral ossification, in situ hybridization analyses indicate that Col10a-1 mRNA are restricted to the hypertrophic regions of growth cartilage.
Persistent Identifierhttp://hdl.handle.net/10722/147384
ISSN
2006 Impact Factor: 3.579
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorKong, RYCen_US
dc.contributor.authorKwan, KMen_US
dc.contributor.authorLau, ETen_US
dc.contributor.authorThomas, JTen_US
dc.contributor.authorBootHandford, RPen_US
dc.contributor.authorGrant, MEen_US
dc.contributor.authorCheah, KSEen_US
dc.date.accessioned2012-05-29T06:03:19Z-
dc.date.available2012-05-29T06:03:19Z-
dc.date.issued1993en_US
dc.identifier.citationEuropean Journal Of Biochemistry, 1993, v. 213 n. 1, p. 99-111en_US
dc.identifier.issn0014-2956en_US
dc.identifier.urihttp://hdl.handle.net/10722/147384-
dc.description.abstractThe entire mouse collagen X gene (Col10a-1) has been isolated. The gene is composed of three exons and two introns spanning 7.0 kb of the DNA sequence. Exons 2 and 3 together encode 15-bp of 5' untranslated sequence, a 2040-bp open reading frame and an 895-nucleotide 3' noncoding region. In the 5' flanking region of the gene, two consensus TATA-box sequences were found. Identification of the first exon by ribonuclease-protection assays and the determination of the 5' end of Col10a-1 mRNA transcripts by primer-extension analyses show that the more 3' TATA box is probably predominantly used and that there are at least three transcription start sites in the exon 1 sequence 3' to this, resulting in 5' untranslated regions of 78, 77 and 55 nucleotides. By means of rapid amplification of cDNA ends by polymerase chain reaction, an additional mRNA species was detected which overlapped the other Col10a-1 transcripts, including the 3' TATA box sequence, giving a 5' untranslated sequence of approximately 235 bases. This layer transcript starts approximately 20 bp 3' to the more 5' TATA box. The data suggest alternative use of promoters and transcription starts for the Col10a-1 gene. Comparison of the combined nucleotide and deduced amino acid sequences of exons 2 and 3 with chicken, bovine and human collagen X genes, showed a high degree of similarity indicating conservation of this gene throughout evolution. Mouse Col10a-1 mRNA was shown to be approximately 3.0 kb and the pepsinized protein. as detected by SDS/PAGE, was ~45 kDa. The mRNA and protein sizes correlate with that predicted by the open reading frame. Reverse-transcription polymerase chain reaction assays indicate that the mouse collagen X gene is first expressed at 13.5 days post coitum, temporally preceding the onset of endochondral ossification. In agreement with the generally accepted association of type-X collagen with endochondral ossification, in situ hybridization analyses indicate that Col10a-1 mRNA are restricted to the hypertrophic regions of growth cartilage.en_US
dc.languageengen_US
dc.publisherBlackwell Publishing Ltd. The Journal's web site is located at http://www.blackwellpublishing.com/journals/EJBen_US
dc.relation.ispartofEuropean Journal of Biochemistryen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshBirdsen_US
dc.subject.meshCartilage - Metabolismen_US
dc.subject.meshCollagen - Genetics - Metabolismen_US
dc.subject.meshCosmidsen_US
dc.subject.meshDnaen_US
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_US
dc.subject.meshExonsen_US
dc.subject.meshFetusen_US
dc.subject.meshGene Expressionen_US
dc.subject.meshHumansen_US
dc.subject.meshIntronsen_US
dc.subject.meshMiceen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPromoter Regions, Geneticen_US
dc.subject.meshRna, Messenger - Metabolismen_US
dc.subject.meshSpecies Specificityen_US
dc.titleIntron-exon structure, alternative use of promoter and expression of the mouse collagen X gene, Col10a-1en_US
dc.typeArticleen_US
dc.identifier.emailCheah, KSE:hrmbdkc@hku.hken_US
dc.identifier.authorityCheah, KSE=rp00342en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid8477738-
dc.identifier.scopuseid_2-s2.0-0027531910en_US
dc.identifier.volume213en_US
dc.identifier.issue1en_US
dc.identifier.spage99en_US
dc.identifier.epage111en_US
dc.identifier.isiWOS:A1993KW05300012-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridKong, RYC=7005290687en_US
dc.identifier.scopusauthoridKwan, KM=7006405800en_US
dc.identifier.scopusauthoridLau, ET=7103086081en_US
dc.identifier.scopusauthoridThomas, JT=7406562866en_US
dc.identifier.scopusauthoridBootHandford, RP=7004054302en_US
dc.identifier.scopusauthoridGrant, ME=7401439700en_US
dc.identifier.scopusauthoridCheah, KSE=35387746200en_US

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