Faculty Research 1990 - 1999

Title

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

Document Type

Article

Publication Date

1996

Keywords

Base-Sequence, Cell-Nucleus, Chromosome-Mapping, DNA: me, DNA-Primers, DNA-Binding-Proteins: me, Germ-Cells, Male, Mice, Mice-Inbred-C57BL, Mice-Transgenic, Molecular-Sequence-Data, Nuclear-Proteins: ge, Promoter-Regions-(Genetics), Regulatory-Sequences-Nucleic-Acid, SUPPORT-U-S-GOVT-NON-P-H-S, SUPPORT-U-S-GOVT-P-H-S, Transcription-Factors: me, Transcription-Genetic

JAX Source

Mol Reprod Dev 1996 Mar;43(3):290-7

Grant

HD24374/HD/NICHD, CA34196/CA/NCI

Abstract

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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