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Originally published In Press as doi:10.1074/jbc.M206065200 on September 3, 2002
J. Biol. Chem., Vol. 277, Issue 45, 43474-43480, November 8, 2002
Human Testis/Sperm-specific Histone H2B (hTSH2B)
MOLECULAR CLONING AND CHARACTERIZATION*
Andrei O.
Zalensky §,
Joseph S.
Siino ,
Arunas A.
Gineitis ,
Irina A.
Zalenskaya ,
Nikolai V.
Tomilin ¶,
Peter
Yau , and
E. Morton
Bradbury
From the Department of Biological Chemistry, School
of Medicine, University of California Davis, Davis, California
95616, ¶ Institute of Cytology, Russian Academy of Sciences, St.
Petersburg 194064, Russia, and Biosciences Division, Los
Alamos National Laboratory, Los Alamos, New Mexico 87545
Human sperm, unlike the sperm of other mammals,
contain replacement histones with unknown biological functions. Here,
we report the identification of the novel human gene coding for a
testis/sperm-specific histone H2B (hTSH2B). This variant histone is
85% homologous to somatic H2B and has over 93% homology with the
testis H2B of rodents. Using genomic PCR, two genetic alleles of hTSH2B
were found in the human population. The hTSH2B gene is transcribed
exclusively in testis, and the corresponding protein is also present in
mature sperm. We expressed recombinant hTSH2B and identified this
protein with a particular H2B subtype expressed in vivo.
The subnuclear distribution of H2B variants in sperm was determined
using biochemical fractionation and immunoblotting. The H2B variant
associated with telomere-binding activity (15) was solubilized by
Triton X-100 or micrococcal nuclease extraction, whereas hTSH2B was
relatively tightly bound in nuclei. Immunofluorescence showed that
hTSH2B was concentrated in spots located at the basal nuclear area of a
subpopulation (20% of cells) of mature sperm. This fact may be of
particular importance, because the hTSH2B "positive" and "negative" sperm cells may undergo significantly different
decondensation processes following fertilization.
*
This work was supported in part by National Institutes of
Health Grant HD39830 (to A. O. Z.) and by Department of Energy Grant FG 0301ER 63070 (to E. M. B.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
§
To whom correspondence should be addressed: Dept. of Biological
Chemistry, School of Medicine, University of California, Davis, CA
95616. Tel.: 530-752-3314; Fax: 530-752-3516; E-mail: aozalensky@ucdavis.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.
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