Biochemical characterization of the Methanothermobacter thermautotrophicus minichromosome maintenance (MCM) helicase N-terminal domains.

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TitleBiochemical characterization of the Methanothermobacter thermautotrophicus minichromosome maintenance (MCM) helicase N-terminal domains.
Publication TypeJournal Article
Year of Publication2004
AuthorsKasiviswanathan, R, Shin, J-H, Melamud, E, Kelman, Z
JournalJ Biol Chem
Volume279
Issue27
Pagination28358-66
Date Published2004 Jul 2
ISSN0021-9258
KeywordsAdenosine Triphosphatases, Archaeal Proteins, Centrifugation, Density Gradient, Chromatography, Gel, DNA Helicases, DNA Replication, DNA, Single-Stranded, Escherichia coli, Genetic Vectors, Glycerol, Kinetics, Methanobacteriaceae, Models, Molecular, Mutation, Protein Binding, Protein Structure, Tertiary, Protein Transport, Software, Streptavidin, Two-Hybrid System Techniques
Abstract

Minichromosome maintenance helicases are ring-shaped complexes that play an essential role in archaeal and eukaryal DNA replication by separating the two strands of chromosomal DNA to provide the single-stranded substrate for the replicative polymerases. For the archaeal protein it was shown that the N-terminal portion of the protein, which is composed of domains A, B, and C, is involved in multimer formation and single-stranded DNA binding and may also play a role in regulating the helicase activity. Here, a detailed biochemical characterization of the N-terminal region of the Methanothermobacter thermautotrophicus minichromosome maintenance helicase is described. Using biochemical and biophysical analyses it is shown that domain C of the N-terminal portion, located adjacent to the helicase catalytic domains, is required for protein multimerization and that domain B is the main contact region with single-stranded DNA. It is also shown that although oligomerization is not essential for single-stranded DNA binding and ATPase activity, the presence of domain C is essential for helicase activity.

DOI10.1074/jbc.M403202200
Alternate JournalJ. Biol. Chem.
PubMed ID15100218