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Abstract
In mammals, the SRY/Sry gene on the Y chromosome is
necessary and sufficient for a bipotential gonad to
develop into testis regardless of its chromosomal sex.
SRY/Sry gene encodes a protein belonging to a high
mobility group (HMG) box protein family and has been
postulated to modulate the expression of genes necessary
for male gonadal differentiation. Using a yeast two-hybrid
screen, we identified a novel protein containing only a
Krüppel associated box (KRAB) domain, which is hereto
named KRAB-O (KRAB Only), as an SRY interacting protein.
KRAB-O protein is encoded by an alternatively spliced
transcript from the Zfp208 locus that also produces
another transcript coding for a KRAB-zinc finger protein,
ZFP208. The interaction of the mouse SRY with KRAB-O was
further confirmed by GST pull-down assay and
co-immunoprecipitation in transfected COS7 cells. The
KRAB-O interaction domain in both the human and mouse SRY
was mapped to the bridge region outside of the HMG box.
Indirect immunofluorescence and confocal microscopy show
that the mouse SRY co-localizes with KRAB-O in nuclear
dots in transiently transfected COS7 cells and primary
fetal mouse gonadal cells. Using similar approaches, we
demonstrate that KRAB-O interacts directly with KAP1 (KRAB
associated protein 1), the obligatory co-repressor for
KRAB domain proteins. Further, we show that the mouse SRY
is associated indirectly with KAP1 and heterochromatin
protein 1 (HP1) through its interaction with KRAB-O,
suggesting that the mouse SRY could utilize the
KRAB-KAP1-HP1 organized transcriptional regulatory complex
to regulate its yet-to-be identified downstream target genes.
Key words:
Testis •
Developmental biology •
Early development •
Male sexual function •
Signal transducers
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