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Originally published In Press as doi:10.1074/mcp.M400102-MCP200 on September 29, 2004.
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Molecular & Cellular Proteomics 3:1170-1180, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Sexual Dimorphism of Rat Liver Nuclear Proteins

Regulatory Role Of Growth Hormone*,S

Ekaterina V. Laz, Christopher A. Wiwi and David J. Waxman{ddagger}

From the Division of Cell and Molecular Biology, Department of Biology, Boston University, Boston, MA 02215

Many genes are expressed in mammalian liver in a sexually dimorphic manner. DNA microarray analysis has shown that growth hormone (GH) and its sex-dependent pattern of pituitary secretion play a major role in establishing the sexually dimorphic patterns of liver gene expression. However, GH may exert effects on protein post-translational modification and nuclear localization that are not reflected at the mRNA level. To investigate these potential effects of GH, we used two-dimensional gel electrophoresis followed by LC-MS/MS to: 1) identify rat liver nuclear proteins whose abundance or state of post-translational modification displays sex-dependent differences; and 2) determine the role of the plasma GH profile in establishing these differences. Nuclear extracts prepared from livers of individual male (n = 9) and female (n = 5) adult rats, and from males given GH by continuous infusion for 7 days to feminize liver gene expression (n = 5 rats), were resolved by two-dimensional electrophoresis. Image analysis of SYPRO Ruby-stained gels revealed 165 sexually dimorphic protein spots that differ in normalized volume between male and female groups by >1.5-fold at p < 0.05. Sixty of these proteins exhibited female-like changes in spot abundance following continuous GH treatment. Comparison of male and GH-treated male groups revealed 130 proteins that displayed >1.5-fold differences in abundance, with 60 of these GH-responsive spots being sexually dimorphic. Thus, GH plays an important role in establishing the sex-dependent differences in liver nuclear protein content. Twenty-eight of the sexually dimorphic and/or GH-regulated protein spots were identified by LC-MS/MS. Proteins identified include regucalcin, nuclear factor 45, and heterogeneous nuclear ribonucleoproteins A3, D-like, and K, in addition to proteins such as GST, normally associated with cytosolic extracts but also reported to be localized in the nucleus.

{ddagger} To whom correspondence should be addressed: Department of Biology, Boston University, 5 Cummington Street, Boston, MA 02215. Fax: 617-353-7404; E-mail: djw{at}bu.edu


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