Large complexes of beta-poly(L-malate) with DNA polymerase alpha, histones, and other proteins in nuclei of growing plasmodia of Physarum polycephalum.
Of the various cell types in the life cycle of Physarum polycephalum, only the growing plasmodium contains the unusual polyester beta-poly(L-malate). The nuclei exhibit large complexes of this polymer with nuclear proteins, among them DNA polymerase alpha, histones, and HMG-like proteins. The complexes are indicated by the results of size exclusion chromatography and chemical cross-linking with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride (EDC). After hydroxylaminolysis of the cross-linked polyester, the proteins are liberated and visualized on Western blots. The complexes of 1200-1400 kDa molecular mass exceed by far the size of free beta-poly(L-malate) and proteins. The observed variation in mass appears to be mainly a function of the kind and stoichiometry of the protein constituents and may explain the relatively high molecular mass in S phase and the low molecular mass during G2 phase of the mitotic cycle. The complexes are considerably stable at moderate ionic strength (100 mM KCl). Also, endogenous beta-poly(L-malate) does not exchange with added beta-[14C]poly(L-malate) during the lysis of the nuclei and the sample preparation. The complexes are dissociated at elevated concentrations of KCl, in the presence of spermine hydrochloride, or by treatment with DEAE/cellulose. Available evidence indicates that beta-poly(L-malate) may be involved in the maintenance of the plasmodial state of P. polycephalum.
Angerer
B
B
Institut für Biophysik und physikalische Biochemie, Universität Regensburg, Germany.
Holler
E
E
D000818:Animals; D002453:Cell Cycle; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002847:Chromatography, Agarose; D003432:Cross-Linking Reagents; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005022:Ethyldimethylaminopropyl Carbodiimide; D006609:High Mobility Group Proteins / Q000378:metabolism; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D008293:Malates / Q000737:chemistry* / Q000378:metabolism; D008938:Mitosis; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D016839:Physarum polycephalum / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding; D013096:Spermine / Q000494:pharmacology
D003432:Cross-Linking Reagents; D006609:High Mobility Group Proteins; D006657:Histones; D006898:Hydroxylamines; D008293:Malates; D009687:Nuclear Proteins; D011108:Polymers; D019811:Hydroxylamine; D013096:Spermine; D011189:Potassium Chloride; C074640:poly(malate); D004257:DNA Polymerase II; D005022:Ethyldimethylaminopropyl Carbodiimide