title: Chromatin binding and polymerization of the endogenous Xenopus egg lamins: the opposing effects of glycogen and ATP.
abstract: We have previously identified and quantitated three B-type lamin isoforms present in the nuclei of mature Xenopus laevis oocytes, and in cell-free egg extracts. As Xenopus egg extracts are frequently used to analyze nuclear envelope assembly and lamina functions, we felt it was imperative that the polymerization and chromatin-binding properties of the endogenous B-type egg lamins be investigated. While we have demonstrated that soluble B-type lamins bind to chromatin, we have also observed that the polymerization of egg lamins does not require membranes or chromatin. Lamin assembly is enhanced by the addition of glycogen/glucose, or by the depletion of ATP from the extract. Moreover, the polymerization of egg cytosol lamins and their binding to demembranated sperm or chromatin assembled from naked lambda-DNA is inhibited by an ATP regeneration system. These ATP-dependent inhibitory activities can be overcome by the coaddition of glycogen to egg cytosol. We have observed that glycogen does not alter ATP levels during cytosol incubation, but rather, as glycogen-enhanced lamin polymerization is inhibited by okadaic acid, we conclude that glycogen activates protein phosphatases. Because protein phosphatase 1 (PP1) is the only phosphatase known to be specifically regulated by glycogen our data indicate that PP1 is involved in lamin polymerization. Our results show that ATP and glycogen effect lamin polymerization and chromatin binding by separate and opposing mechanisms.
authors.affiliation: Division of Electron Microscopy, Biocenter of the University of W urzburg, Am Hubland, D-97074 Würzburg, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Lourim
authors.forename: G
authors.initials: G
authors.lastname: Krohne
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001704:Biopolymers / Q000096:biosynthesis / Q000737:chemistry; D002457:Cell Extracts / Q000737:chemistry; D002843:Chromatin / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D005260:Female; D006003:Glycogen / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000378:metabolism; D034882:Lamins; D008297:Male; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D009865:Oocytes / Q000737:chemistry; D011485:Protein Binding / Q000187:drug effects; D020033:Protein Isoforms / Q000378:metabolism; D013347:Subcellular Fractions / Q000737:chemistry; D014982:Xenopus laevis
chemical_list: D001704:Biopolymers; D002457:Cell Extracts; D002843:Chromatin; D034882:Lamins; D009687:Nuclear Proteins; D020033:Protein Isoforms; D000255:Adenosine Triphosphate; D006003:Glycogen