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Binding of RFX2 and NF‐Y to the testis‐specific histone H1t promoter may be required for transcriptional activation in primary spermatocytes
Author(s) -
vanWert Jane M.,
Wolfe Steven A.,
Grimes Sidney R.
Publication year - 2008
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.21694
Subject(s) - caat box , microbiology and biotechnology , promoter , biology , transcription factor , histone h1 , histone , gene expression , gene , genetics
The testis‐specific linker histone H1t is transcribed exclusively in pachytene spermatocytes during spermatogenesis. The H1t promoter contains two imperfect inverted repeats which together comprise the X‐box motif that is known to bind the transcription factor regulatory factor X (RFX). Out of all the histone H1 family promoters this motif appears only in the H1t promoter and may contribute to H1t tissue‐specific expression. We show by Western blotting, EMSA, ChIP analysis, and real‐time RT‐PCR that the rat H1t X‐box is bound by RFX2 in vivo in spermatocytes. We demonstrate that transcription factor NF‐Y binds to the CCAAT‐box motif that is located downstream and adjacent to the X‐box and that testis NF‐Y interacts either directly or indirectly with RFX2. Furthermore, we show that both the X‐box and CCAAT‐box are required for promoter activity and that co‐expression of RFX2 greatly enhances testis histone H1t promoter activity in the GC‐1spg germinal cell line. J. Cell. Biochem. 104: 1087–1101, 2008. © 2008 Wiley‐Liss, Inc.