TY - JOUR
T1 - Pairwise interactions of the six human MCM protein subunits
AU - Yu, Zhiling
AU - Feng, Daorong
AU - Liang, Chun
N1 - Funding Information:
We thank Cathy Shum, Mandy Chan, Kevin Ng, Tony Suen and Kelvin Sou for assistance. This work was supported by the Hong Kong Research Grants Council.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2004/7/23
Y1 - 2004/7/23
N2 - The eukaryotic minichromosome maintenance (MCM) proteins have six subunits, Mcm2 to 7p. Together they play essential roles in the initiation and elongation of DNA replication, and the human MCM proteins present attractive targets for potential anticancer drugs. The six MCM subunits interact and form a ring-shaped heterohexameric complex containing one of each subunit in a variety of eukaryotes, and subcomplexes have also been observed. However, the architecture of the human MCM heterohexameric complex is still unknown. We systematically studied pairwise interactions of individual human MCM subunits by using the yeast two-hybrid system and in vivo protein-protein crosslinking with a non-cleavable crosslinker in human cells followed by co-immunoprecipitation. In the yeast two-hybrid assays, we revealed multiple binary interactions among the six human MCM proteins, and a subset of these interactions was also detected as direct interactions in human cells. Based on our results, we propose a model for the architecture of the human MCM protein heterohexameric complex. We also propose models for the structures of subcomplexes. Thus, this study may serve as a foundation for understanding the overall architecture and function of eukaryotic MCM protein complexes and as clues for developing anticancer drugs targeted to the human MCM proteins.
AB - The eukaryotic minichromosome maintenance (MCM) proteins have six subunits, Mcm2 to 7p. Together they play essential roles in the initiation and elongation of DNA replication, and the human MCM proteins present attractive targets for potential anticancer drugs. The six MCM subunits interact and form a ring-shaped heterohexameric complex containing one of each subunit in a variety of eukaryotes, and subcomplexes have also been observed. However, the architecture of the human MCM heterohexameric complex is still unknown. We systematically studied pairwise interactions of individual human MCM subunits by using the yeast two-hybrid system and in vivo protein-protein crosslinking with a non-cleavable crosslinker in human cells followed by co-immunoprecipitation. In the yeast two-hybrid assays, we revealed multiple binary interactions among the six human MCM proteins, and a subset of these interactions was also detected as direct interactions in human cells. Based on our results, we propose a model for the architecture of the human MCM protein heterohexameric complex. We also propose models for the structures of subcomplexes. Thus, this study may serve as a foundation for understanding the overall architecture and function of eukaryotic MCM protein complexes and as clues for developing anticancer drugs targeted to the human MCM proteins.
KW - GST, glutathione-S-transferase
KW - MCM, minichromosome maintenance
KW - SCM, synthetic complete medium
KW - WCE, whole cell extract
KW - co-immunoprecipitation
KW - human MCM proteins
KW - protein-protein crosslinking
KW - protein-protein interaction
KW - yeast two-hybrid system
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U2 - 10.1016/j.jmb.2004.05.024
DO - 10.1016/j.jmb.2004.05.024
M3 - Article
C2 - 15236977
AN - SCOPUS:3242770626
SN - 0022-2836
VL - 340
SP - 1197
EP - 1206
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 5
ER -