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Title: MCM ring hexamerization is a prerequisite for DNA-binding

Abstract

The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in the hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.

Authors:
 [1];  [1];  [1]
  1. St. Jude Children's Research Hospital, Memphis, TN (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1223715
Grant/Contract Number:  
AC02-06CH11357; W-31-109-ENG-38
Resource Type:
Accepted Manuscript
Journal Name:
Nucleic Acids Research
Additional Journal Information:
Journal Name: Nucleic Acids Research; Journal ID: ISSN 0305-1048
Publisher:
Oxford University Press
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Froelich, Clifford A., Nourse, Amanda, and Enemark, Eric J. MCM ring hexamerization is a prerequisite for DNA-binding. United States: N. p., 2015. Web. doi:10.1093/nar/gkv914.
Froelich, Clifford A., Nourse, Amanda, & Enemark, Eric J. MCM ring hexamerization is a prerequisite for DNA-binding. United States. doi:10.1093/nar/gkv914.
Froelich, Clifford A., Nourse, Amanda, and Enemark, Eric J. Sun . "MCM ring hexamerization is a prerequisite for DNA-binding". United States. doi:10.1093/nar/gkv914. https://www.osti.gov/servlets/purl/1223715.
@article{osti_1223715,
title = {MCM ring hexamerization is a prerequisite for DNA-binding},
author = {Froelich, Clifford A. and Nourse, Amanda and Enemark, Eric J.},
abstractNote = {The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in the hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.},
doi = {10.1093/nar/gkv914},
journal = {Nucleic Acids Research},
number = ,
volume = ,
place = {United States},
year = {2015},
month = {9}
}

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