Kohn, M.J., K.J. Kaneko and M.L. DePamphilis (2005) DkkL1(Soggy), A Dickkopf Family Member, Localizes To The Acrosome During Mammalian Spermatogenesis, Mol. Reprod. Dev. 71:516-522. Dickkopf-like 1 (DkkL1) is related to the Dickkopf gene family, a group of proteins that are characterized as secreted antagonists of Wingless signal transduction proteins. DkkL1 mRNA is found in preimplantation mouse embryos and in developing neural tissue, but in adults it is found primarily in the testes. In an effort to elucidate its function, the distribution of Dickkopf-like 1 protein (DkkL1) in mouse testis and mature sperm was analyzed by immuno-histochemistry and immuno-blotting techniques. DkkL1 first appeared in the developing spermatocytes in seminiferous tubules as early as Stage XII, coincident with the appearance of DkkL1 mRNA. Surprisingly, however, DkkL1 localized to the developing acrosome in spermatocytes and spermatids and to the acrosome in mature sperm. Furthermore, DkkL1 was N-glycosylated in the testis, but it did not appear to be excreted, and the DkkL1 in mature sperm was no longer N-glycosylated, suggesting that additional post-translational modifications occurred during the final stages of spermatogenesis. These results identify a member of the Dickkopf family as a novel acrosomal protein that may be involved in acrosome assembly or function, a unique role for a secreted signaling molecule. Kaneko, K.J., T. Rein, Z-S Guo, K. Latham and M.L. DePamphilis. (2004) DNA Methylation May Restrict But Does Not Determine Differential Gene Expression at the Sgy/Tead2 locus During Mouse Development, Mol. Cell. Biol. 24:1968-1982. Soggy (Sgy) and Tead2, two closely linked genes with CpG islands, were coordinately expressed in mouse preimplantation embryos and embryonic stem (ES) cells, but were differentially expressed in differentiated cells. Analysis of established cell lines revealed that Sgy gene expression could be fully repressed by methylation of the Sgy promoter, and that DNA methylation acted synergistically with chromatin deacetylation. Differential gene expression correlated with differential DNA methylation, resulting in sharp transitions from methylated to unmethylated DNA at the open promoter in both normal cells and tissues, as well as in established cell lines. However, neither promoter was methylated in normal cells and tissues even when its transcripts were undetectable. Moreover, the Sgy promoter remained unmethylated as Sgy expression was repressed during ES cell differentiation. Therefore, DNA methylation was not the primary determinant of Sgy/Tead2 expression. Nevertheless, Sgy expression was consistently restricted to basal levels whenever downstream regulatory sequences were methylated, suggesting that DNA methylation restricts but does not regulate differential gene expression during mouse development. Intine, R. V., M. Dundr, A. Vassilev, E. Schwartz, Y. Zhou, Y. Zhao, M. L. DePamphilis, and R. J. Maraia (2004) Nonphosphorylated Human La Antigen Interacts With Nucleolin At Nucleolar Sites Involved In rRNA Biogenesis, Mol. Cell. Biol. 24:10894-904. La is an abundant RNA-binding protein that has been proposed to function in multiple pathways involved in the production of tRNAs, ribosomal proteins and other components of the translational machinery (Kenan and Keene, 2004 Nat Struct & Mol Biol, 11:306). Human (h)La is either phosphorylated or nonphosphorylated on serine-366 (S366, pLa & npLa, respectively). In addition to its conserved N-terminal domain, hLa contains a C-terminal domain that contains an atypical RNA recognition motif (RRM) and short basic motif (SBM), whose distinct mode of RNA binding is inhibited by S366 phosphorylation. We report that npLa is most concentrated in nucleolar foci that harbor fibrillarin, nucleolin, and nascent pol I transcripts. Immunoprecipitation of native or tagged hLa as well as yeast two hybrid analysis and fluorescence resonance energy transfer (FRET) in the nucleolus, reveals association with nucleolin. HLa lacking the SBM fails to localize to nucleoli and does not exhibit FRET with nucleolin. The data indicate that the nonphosphorylated SBM directs interaction with nucleolin, and provides evidence that the npLa specifically is involved in nucleolar biology related to ribosome biogenesis. DePamphilis, M.L. (2004) Mammalian Development, Regulation of Gene Expression in, in Meyers, R.A. (Ed.): Encyclopedia of Molecular Cell Biology and Molecular Medicine, volume 6, pp. ??, WILEY-VCH Verlag, Weinheim, Germany. (www.wiley-vch.de/books/emcbmm2/index.html)

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U.S. National Inst/Child Hlth/Human Dev
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Ullah, Zakir; Lee, Chrissie Y; Depamphilis, Melvin L (2009) Cip/Kip cyclin-dependent protein kinase inhibitors and the road to polyploidy. Cell Div 4:10
Yagi, Rieko; Kohn, Matthew J; Karavanova, Irina et al. (2007) Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development. Development 134:3827-36
Kaneko, Kotaro J; Kohn, Matthew J; Liu, Chengyu et al. (2007) Transcription factor TEAD2 is involved in neural tube closure. Genesis 45:577-87
Park, Jung-Min; Kohn, Matthew J; Bruinsma, Monique W et al. (2006) The multifunctional RNA-binding protein La is required for mouse development and for the establishment of embryonic stem cells. Mol Cell Biol 26:1445-51
Kohn, Matthew J; Kaneko, Kotaro J; DePamphilis, Melvin L (2005) DkkL1 (Soggy), a Dickkopf family member, localizes to the acrosome during mammalian spermatogenesis. Mol Reprod Dev 71:516-22
Kaneko, Kotaro J; Rein, Theo; Guo, Zong-Sheng et al. (2004) DNA methylation may restrict but does not determine differential gene expression at the Sgy/Tead2 locus during mouse development. Mol Cell Biol 24:1968-82
Intine, Robert V; Dundr, Miroslav; Vassilev, Alex et al. (2004) Nonphosphorylated human La antigen interacts with nucleolin at nucleolar sites involved in rRNA biogenesis. Mol Cell Biol 24:10894-904