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PMID: 10606645 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Nucleoplasmin-mediated chromatin remodelling is required for Xenopus sperm nuclei to become licensed for DNA replication.

Nucleic acids research ·Vol. 28 ·No. 2 ·2000-01-15 ·Pages 472-80

Gillespie PJ, Blow JJ

Abstract

During late mitosis and early G(1), a series of proteins are assembled onto replication origins, resulting in them becoming 'licensed' for replication in the subsequent S phase. Four factors have so far been identified that are required for chromatin to become functionally licensed: ORC (the origin recognition complex) and Cdc6, plus the two components of the replication licensing system RLF-M and RLF-B. Here we describe the first steps of a systematic fractionation of Xenopus egg extracts to identify all the components necessary for the assembly of licensed replication origins on Xenopus sperm nuclei (the physiological DNA substrate in this system). We have purified a new activity essential for this reaction, and have shown that it is nucleoplasmin, a previously known chromatin remodelling protein. Nucleoplasmin decondenses the sperm chromatin by removing protamines, and is required at the earliest known step in origin assembly to allow ORC to bind to the DNA. Sperm nuclei can be licensed by a combination of nucleoplasmin, RLF-M and a partially purified fraction that contains ORC, Cdc6 and RLF-B. This suggests that we are likely to have identified most of the proteins required for this assembly reaction.

MeSH Terms
Animals Cell Nucleus/metabolism Chromatin/metabolism DNA/metabolism DNA Replication DNA-Binding Proteins/isolation & purification,metabolism Male Nuclear Proteins/physiology Nucleoplasmins Origin Recognition Complex Phosphoproteins/physiology Protein Binding Spermatozoa/metabolism Xenopus
Chemicals
Chromatin DNA-Binding Proteins Nuclear Proteins Nucleoplasmins Origin Recognition Complex Phosphoproteins DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gillespie P J
CRC Chromosome Replication Research Group, Division of Gene Regulation,Department of Biochemistry, University of Dundee, Dundee DD1 5EH, UK.
Blow J J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/28.2.472
Published
2000-01-15
Pages
472-80
Language
English
Region
England
NLM ID
0411011
PMCID
PMC102529
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/28.2.472 PMC Full Text View journal details
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