Pre-replicative association of multiple replicative enzyme activities with the nuclear matrix during rat liver regeneration.
As a step toward the molecular elucidation of the putative replicational apparatus associated with the nuclear matrix, we have investigated the possible matrix association of several replicational related enzymes. In addition to the previously identified DNA polymerase alpha, DNA primase, 3'-5' exonuclease, RNase H, and DNA methylase were all recovered at significant levels (20-30% of total nuclear activity) in nuclear matrix isolated from regenerating rat liver during maximal in vivo replication (22 h post-hepatectomy). In contrast, DNA ligase was not detected on the nuclear matrix even though significant activity was present in isolated nuclei. Examination of the replicative dependency of these enzyme activities following partial hepatectomy revealed pre-replicative elevations which were distinct for each matrix-bound enzyme. A second late-replicative peak in DNA methylase is consistent with a role of this matrix-bound enzyme in the maintenance of the inheritable methylation pattern. Mild sonication resulted in a significant release of all of these activities except RNase H. A major portion of the matrix-solubilized DNA polymerase alpha, DNA primase, 3'-5' exonuclease, and DNA methylase activities cosedimented on sucrose gradients between approximately 8-12 S. Our results are consistent with the organization of at least a portion of these replicative enzymes into nuclear matrix-bound replicational complexes. We also propose a novel pre-replicative assembly model of the matrix-bound replicational apparatus in which DNA primase plays an initial and critical role.
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D004257:DNA Polymerase II / Q000378:metabolism; D019915:DNA Primase; D004261:DNA Replication; D004722:Endoribonucleases / Q000378:metabolism; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism; D006498:Hepatectomy; D008099:Liver / Q000648:ultrastructure*; D008115:Liver Regeneration; D012316:RNA Nucleotidyltransferases / Q000378:metabolism; D051381:Rats; D016914:Ribonuclease H
D004248:DNA (Cytosine-5-)-Methyltransferases; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II; D004722:Endoribonucleases; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; D016914:Ribonuclease H