cAMP-responsive element in TATA-less core promoter is essential for haploid-specific gene expression in mouse testis.
Promoters, including neither TATA box nor initiator, have been frequently found in testicular germ cell-specific genes in mice. These investigations imply that unique forms of the polymerase II transcription initiation machinery play a role in selective activation of germ cell-specific gene expression programs during spermatogenesis. However, there is little information about testis-specific core promoters, because useful germ cell culture system is not available. In this study, we characterize the regulatory region of the haploid-specific Oxct2b gene in detail by using in vivo transient transfection assay in combination with a transgenic approach, with electrophoretic mobility shift and chromatin immunoprecipitation assays. Expression studies using mutant constructs demonstrate that a 34 bp region, which extends from -49 to -16, acts as a core promoter in an orientation-dependent manner. This promoter region includes the cAMP-responsive element (CRE)-like sequence TGACGCAG, but contains no other motifs, such as a TATA box or initiator. The CRE-like element is indispensable for the core promoter activity, but not for activator in testicular germ cells, through the binding of a testis-specific CRE modulator transcription factor. These results indicate the presence of alternative transcriptional initiation machinery for cell-type-specific gene expression in testicular germ cells.
Somboonthum
Pranee
P
Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University Suita, Osaka 565-0871, Japan.
Ohta
Hiroshi
H
Yamada
Shuichi
S
Onishi
Masayoshi
M
Ike
Akiko
A
Nishimune
Yoshitake
Y
Nozaki
Masami
M
D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015256:Coenzyme A-Transferases / Q000096:biosynthesis / Q000235:genetics*; D000242:Cyclic AMP / Q000378:metabolism; D051786:Cyclic AMP Response Element Modulator; D004268:DNA-Binding Proteins / Q000378:metabolism*; D018274:Electroporation; D006238:Haploidy; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D020218:Response Elements; D016415:Sequence Alignment; D013087:Spermatids / Q000378:metabolism; D016385:TATA Box; D013737:Testis / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
D004268:DNA-Binding Proteins; D014157:Transcription Factors; D051786:Cyclic AMP Response Element Modulator; D000242:Cyclic AMP; D015256:Coenzyme A-Transferases; C021460:3-ketoacid CoA-transferase