Cell cycle-dependent expression of nuclear matrix proteins of Ehrlich ascites cells studied by in vitro translation.
A combination of methods was used to study the cell cycle-dependent expression of nuclear matrix proteins of Ehrlich ascites cells: Separation of asynchronous cells growing in vivo into fractions of G1-, S- and G2- phase cells by centrifugal elutriation with less than 10% cross-contamination. Isolation of poly(A+) RNA populations from total cytoplasmic RNA by affinity chromatography on messenger affinity paper (mAP). In vitro translation of poly(A+) RNA from asynchronous and phase synchronous cells. Immunoprecipitation of in vitro synthesized nuclear matrix proteins by a monoclonal antibody with anti-lamin specificity (PKB8) and by a polyspecific anti-nuclear matrix serum (AMS5) followed by analysis of immunoprecipitated materials on SDS-polyacrylamide gels. The results indicate that mRNAs for nuclear matrix-associated proteins including the lamins B and C are either exclusively or at least predominantly present in the cytoplasm of cells in S phase suggesting a high rate of in vivo synthesis of these proteins during S phase. This is consistent with an anticipated biological function of the nuclear matrix which is considered to organize parental and newly synthesized DNA in higher order structures.
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Kopun
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Werner
D000818:Animals; D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D002453:Cell Cycle; D002458:Cell Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D009698:Nucleoproteins / Q000096:biosynthesis*; D011061:Poly A / Q000378:metabolism; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D009698:Nucleoproteins; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA