TY - JOUR
T1 - Human DNA helicase B interacts with the replication initiation protein Cdc45 and facilitates Cdc45 binding onto chromatin
AU - Gerhardt, Jeannine
AU - Guler, Gulfem D.
AU - Fanning, Ellen
N1 - Funding Information:
We thank D. Cortez, A. Dutta, F. Haenel, X. Jiang, W. C. Lin, A. Rokas, D. Schaarschmidt, B. Stillman and E. Warren for reagents and discussion. JG, GG and EF designed experiments, JG and GG performed experiments, JG and GG analyzed data and wrote the manuscript. The financial support of NIH ( GM52948 to EF and P30 CA068485 to the Vanderbilt-Ingram Cancer Center ), Helmholtz Center Munich, and Vanderbilt University are gratefully acknowledged.
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/6/10
Y1 - 2015/6/10
N2 - The chromosomal DNA replication in eukaryotic cells begins at replication initation sites, which are marked by the assembly of the pre-replication complexes in early G1. At the G1/S transition, recruitment of additional replication initiation proteins enables origin DNA unwinding and loading of DNA polymerases. We found that depletion of the human DNA helicase B (HDHB) inhibits the initiation of DNA replication, suggesting a role of HDHB in the beginning of the DNA synthesis. To gain insight into the function of HDHB during replication initiation, we examined the physical interactions of purified recombinant HDHB with key initiation proteins. HDHB interacts directly with two initiation factors TopBP1 and Cdc45. In addition we found that both, the N-terminus and helicase domain of HDHB bind to the N-terminus of Cdc45. Furthermore depletion of HDHB from human cells diminishes Cdc45 association with chromatin, suggesting that HDHB may facilitate Cdc45 recruitment at G1/S in human cells.
AB - The chromosomal DNA replication in eukaryotic cells begins at replication initation sites, which are marked by the assembly of the pre-replication complexes in early G1. At the G1/S transition, recruitment of additional replication initiation proteins enables origin DNA unwinding and loading of DNA polymerases. We found that depletion of the human DNA helicase B (HDHB) inhibits the initiation of DNA replication, suggesting a role of HDHB in the beginning of the DNA synthesis. To gain insight into the function of HDHB during replication initiation, we examined the physical interactions of purified recombinant HDHB with key initiation proteins. HDHB interacts directly with two initiation factors TopBP1 and Cdc45. In addition we found that both, the N-terminus and helicase domain of HDHB bind to the N-terminus of Cdc45. Furthermore depletion of HDHB from human cells diminishes Cdc45 association with chromatin, suggesting that HDHB may facilitate Cdc45 recruitment at G1/S in human cells.
KW - Cdc45
KW - DNA replication initiation
KW - Human DNA Helicase B
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U2 - 10.1016/j.yexcr.2015.04.014
DO - 10.1016/j.yexcr.2015.04.014
M3 - Article
C2 - 25933514
AN - SCOPUS:84930199084
VL - 334
SP - 283
EP - 293
JO - Experimental Cell Research
JF - Experimental Cell Research
SN - 0014-4827
IS - 2
ER -