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Mammalian spermiogenesis is marked by the initial disruption of
the nuclear-histone-DNA complex by the transition proteins for ultimate
replacement with protamines. The genes for three of these low molecular
weight basic nuclear proteins exist as a single linear array of
PRM1, PRM2, and TNP2 on human chromosome
16p13.2. To begin to address the mechanism governing their
transcriptional potentiation, a region of
Volume 270,
Number 15,
Issue of April 14, pp. 8755-8762, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
40 kilobases of the
human genome encompassing these genes was introduced into the germ line
of mice. Fluorescence in situ hybridization and Southern
analysis showed that this segment of the human genome integrated into
independent chromosomal sites while maintaining its fidelity.
Transcript analysis demonstrated that the expression of the endogenous
mouse protamine Prm1 and Prm2 genes as well as the
mouse transition protein Tnp2 gene were expressed along with
their human transgene counterparts. The pattern of expression of these
transgenic human genes within this multigenic cluster faithfully
represented that observed in vivo. In addition, all members of
this transgenic gene cluster were expressed in proportions similar to
those in human testis. Copy number-dependent and position-independent
expression of the transgenic construct demonstrated that the
corresponding biological locus was contained within this segment of the
human genome. Furthermore, DNase I sensitivity established that in
sperm the human PRM1
PRM2
TNP2 genic domain was contained as an
28.5-kilobase contiguous
segment bounded by an array of nuclear matrix associated topoisomerase
II consensus sites. This is the first description of a multigenic male
gamete-specific domain as a fundamental gene regulatory unit. A model
of haploid-specific gene determination is presented.
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