Crystal structure of the human Polϵ B-subunit in complex with the C-terminal domain of the catalytic subunit

Baranovskiy, Andrey G.; Gu, Jianyou; Babayeva, Nigar D.; Kurinov, Igor; Pavlov, Youri I.; Tahirov, Tahir H.

doi:10.1074/jbc.M117.792705

Title: Crystal structure of the human Polϵ B-subunit in complex with the C-terminal domain of the catalytic subunit

Journal Article · Fri Sep 01 00:00:00 EDT 2017 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.M117.792705· OSTI ID:1769198

;

; Babayeva, Nigar D.; Kurinov, Igor; Pavlov, Youri I.;

The eukaryotic B-family DNA polymerases include four members: Pol$$α$$, Pol$$δ$$, Pol$$ϵ$$, and Pol$$ζ$$, which share common architectural features, such as the exonuclease/polymerase and C-terminal domains (CTDs) of catalytic subunits bound to indispensable B-subunits, which serve as scaffolds that mediate interactions with other components of the replication machinery. Crystal structures for the B-subunits of Pol$$α$$ and Pol$$δ$$/Pol$$ζ$$ have been reported: the former within the primosome and separately with CTD and the latter with the N-terminal domain of the C-subunit. In this paper we present the crystal structure of the human Pol$$ϵ$$ B-subunit (p59) in complex with CTD of the catalytic subunit (p261_C). The structure revealed a well defined electron density for p261_C and the phosphodiesterase and oligonucleotide/oligosaccharide-binding domains of p59. However, electron density was missing for the p59 N-terminal domain and for the linker connecting it to the phosphodiesterase domain. Similar to Polα, p261_C of Pol$$ϵ$$ contains a three-helix bundle in the middle and zinc-binding modules on each side. Intersubunit interactions involving 11 hydrogen bonds and numerous hydrophobic contacts account for stable complex formation with a buried surface area of 3094 Å². Comparative structural analysis of p59–p261_C with the corresponding Polα complex revealed significant differences between the B-subunits and CTDs, as well as their interaction interfaces. The B-subunit of Pol$$δ$$/Pol$$ζ$$ also substantially differs from B-subunits of either Pol$$α$$ or Pol$$ϵ$$. This work provides a structural basis to explain biochemical and genetic data on the importance of B-subunit integrity in replisome function in vivo.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); USDOE Office of Science (SC)

Grant/Contract Number:: AC02–06CH11357; P41 GM103403; S10 RR029205; AC02-06CH11357

OSTI ID:: 1769198

Alternate ID(s):: OSTI ID: 1600812

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 292 Journal Issue: 38; ISSN 0021-9258

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 25 works

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
crystal structure
DNA replication
human
protein complex
zinc
B-subunit
DNA polymerase epsilon
Dpb2
zinc-binding module

Title: Crystal structure of the human Polϵ B-subunit in complex with the C-terminal domain of the catalytic subunit

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