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Article: Mouse testin: Complementary DNA cloning, genomic organization, and characterization of its proximal promoter region

TitleMouse testin: Complementary DNA cloning, genomic organization, and characterization of its proximal promoter region
Authors
KeywordsEarly development
Gene regulation
Sertoli cells
Spermatogenesis
Testis
Issue Date2003
PublisherSociety for the Study of Reproduction. The Journal's web site is located at http://www.biolreprod.org/
Citation
Biology Of Reproduction, 2003, v. 68 n. 4, p. 1376-1386 How to Cite?
AbstractTestin is a secretory protein that was initially identified from rat Sertoli cell-enriched cultures and has been suggested to be a sensitive marker to monitor the integrity of Sertoli-germ cell junctions. However, the expression of the testin gene in other species and the molecular mechanisms that govern its transcription are unknown. To address these issues, we cloned and characterized the mouse testin gene. A full-length mouse testin cDNA encoding a polypeptide of 333 amino acid residues was isolated by library screening. Sequence analysis revealed that mouse testin shares 90.1%, 58.9%, 62.2%, and 64.6% identity with rat testin and cathepsin L of mouse, rat, and human, respectively, at the amino acid level. Reverse transcription-polymerase chain reaction and Southern blot analysis demonstrated that mouse testin transcripts were predominantly expressed in the gonads. The mouse testin gene spans over 21 kilobases (kb) and contains eight exons interrupted by seven introns. Primer extension analysis and 5′ rapid amplification of cDNA ends identified a major transcription start site located 134 base pairs upstream from the translation initiation codon. Analysis of a 2.3-kb mouse testin 5′-flanking region revealed that it lacked TATA and CAAT boxes, and the region was not GC rich. By the use of deletion analysis, in vitro DNase I footprinting, and site-directed mutagenesis, we identified within the proximal promoter region three closely spaced putative binding sites for GATA, sex-determining factor, and steroidogenic factor 1 that are important for testin gene transcription in mouse Sertoli (MSC-1) cells. These cis-acting elements are also present in the conserved Mullerian-inhibiting substance (MIS) proximal promoters, raising a possibility that the transcriptions of testin and MIS genes are controlled by similar mechanisms.
Persistent Identifierhttp://hdl.handle.net/10722/84872
ISSN
2015 Impact Factor: 3.471
2015 SCImago Journal Rankings: 1.646
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChi, KCen_HK
dc.contributor.authorChiu, HCen_HK
dc.contributor.authorLee, WMen_HK
dc.date.accessioned2010-09-06T08:58:06Z-
dc.date.available2010-09-06T08:58:06Z-
dc.date.issued2003en_HK
dc.identifier.citationBiology Of Reproduction, 2003, v. 68 n. 4, p. 1376-1386en_HK
dc.identifier.issn0006-3363en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84872-
dc.description.abstractTestin is a secretory protein that was initially identified from rat Sertoli cell-enriched cultures and has been suggested to be a sensitive marker to monitor the integrity of Sertoli-germ cell junctions. However, the expression of the testin gene in other species and the molecular mechanisms that govern its transcription are unknown. To address these issues, we cloned and characterized the mouse testin gene. A full-length mouse testin cDNA encoding a polypeptide of 333 amino acid residues was isolated by library screening. Sequence analysis revealed that mouse testin shares 90.1%, 58.9%, 62.2%, and 64.6% identity with rat testin and cathepsin L of mouse, rat, and human, respectively, at the amino acid level. Reverse transcription-polymerase chain reaction and Southern blot analysis demonstrated that mouse testin transcripts were predominantly expressed in the gonads. The mouse testin gene spans over 21 kilobases (kb) and contains eight exons interrupted by seven introns. Primer extension analysis and 5′ rapid amplification of cDNA ends identified a major transcription start site located 134 base pairs upstream from the translation initiation codon. Analysis of a 2.3-kb mouse testin 5′-flanking region revealed that it lacked TATA and CAAT boxes, and the region was not GC rich. By the use of deletion analysis, in vitro DNase I footprinting, and site-directed mutagenesis, we identified within the proximal promoter region three closely spaced putative binding sites for GATA, sex-determining factor, and steroidogenic factor 1 that are important for testin gene transcription in mouse Sertoli (MSC-1) cells. These cis-acting elements are also present in the conserved Mullerian-inhibiting substance (MIS) proximal promoters, raising a possibility that the transcriptions of testin and MIS genes are controlled by similar mechanisms.en_HK
dc.languageengen_HK
dc.publisherSociety for the Study of Reproduction. The Journal's web site is located at http://www.biolreprod.org/en_HK
dc.relation.ispartofBiology of Reproductionen_HK
dc.subjectEarly developmenten_HK
dc.subjectGene regulationen_HK
dc.subjectSertoli cellsen_HK
dc.subjectSpermatogenesisen_HK
dc.subjectTestisen_HK
dc.titleMouse testin: Complementary DNA cloning, genomic organization, and characterization of its proximal promoter regionen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0006-3363&volume=68&spage=1376&epage=1386&date=2003&atitle=Mouse+Testin:+Complementary+DNA+Cloning,+Genomic+Organization,+and+Characterization+of+Its+Proximal+Promoter+Regionen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1095/biolreprod.102.011205en_HK
dc.identifier.pmid12606342en_HK
dc.identifier.scopuseid_2-s2.0-0037379493en_HK
dc.identifier.hkuros81139en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037379493&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume68en_HK
dc.identifier.issue4en_HK
dc.identifier.spage1376en_HK
dc.identifier.epage1386en_HK
dc.identifier.isiWOS:000181844400038-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridChi, KC=7102221552en_HK
dc.identifier.scopusauthoridChiu, HC=7401986734en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK

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