8868241

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3

Transgenic mice were generated to localize essential promoter elements in the mouse testis-expressed Tcp-10 genes. These genes are expressed exclusively in male germ cells, and exhibit a diffuse range of transcriptional start sites, possibly due to the absence of a TATA box. A series of transgene constructs containing different amounts of 5' flanking DNA revealed that all sequences necessary for appropriate temporal and tissue-specific transcription of Tcp-10 reside between positions -1 to -973. All transgenic animals containing these sequences expressed a chimeric transgene at high levels, in a pattern that paralleled the endogenous genes. These experiments further defined a 227 bp fragment from -746 to -973 that was absolutely essential for expression. In a gel-shift assay, this 227-bp fragment bound nuclear protein from testis, but not other tissues, to yield two retarded bands. Sequence analysis of this fragment revealed a half-site for the AP-2 transcription factor recognition sequence. Gel shift assays using native or mutant oligonucleotides demonstrated that the putative AP-2 recognition sequence was essential for generating the retarded bands. Since the binding activity is testis-specific, but AP-2 expression is not exclusive to male germ cells, it is possible that transcription of Tcp-10 requires interaction between AP-2 and a germ cell-specific transcription factor.

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1040-452X

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290-7

pubmed-meshheading:8868241-Animals, pubmed-meshheading:8868241-Base Sequence, pubmed-meshheading:8868241-Cell Nucleus, pubmed-meshheading:8868241-Chromosome Mapping, pubmed-meshheading:8868241-DNA, pubmed-meshheading:8868241-DNA Primers, pubmed-meshheading:8868241-DNA-Binding Proteins, pubmed-meshheading:8868241-Germ Cells, pubmed-meshheading:8868241-Intracellular Signaling Peptides and Proteins, pubmed-meshheading:8868241-Male, pubmed-meshheading:8868241-Mice, pubmed-meshheading:8868241-Mice, Inbred C57BL, pubmed-meshheading:8868241-Mice, Transgenic, pubmed-meshheading:8868241-Microtubule-Associated Proteins, pubmed-meshheading:8868241-Molecular Sequence Data, pubmed-meshheading:8868241-Nuclear Proteins, pubmed-meshheading:8868241-Promoter Regions, Genetic, pubmed-meshheading:8868241-Proteins, pubmed-meshheading:8868241-Regulatory Sequences, Nucleic Acid, pubmed-meshheading:8868241-Transcription, Genetic, pubmed-meshheading:8868241-Transcription Factor AP-2, pubmed-meshheading:8868241-Transcription Factors, pubmed-meshheading:8868241-t-Complex Genome Region

Promoter mapping of the mouse Tcp-10bt gene in transgenic mice identifies essential male germ cell regulatory sequences.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S.