title: Crystal Structure of the Human Pol α B Subunit in Complex with the C-terminal Domain of the Catalytic Subunit.
abstract: In eukaryotic DNA replication, short RNA-DNA hybrid primers synthesized by primase-DNA polymerase α (Prim-Pol α) are needed to start DNA replication by the replicative DNA polymerases, Pol δ and Pol ϵ. The C terminus of the Pol α catalytic subunit (p180C) in complex with the B subunit (p70) regulates the RNA priming and DNA polymerizing activities of Prim-Pol α. It tethers Pol α and primase, facilitating RNA primer handover from primase to Pol α. To understand these regulatory mechanisms and to reveal the details of human Pol α organization, we determined the crystal structure of p70 in complex with p180C. The structured portion of p70 includes a phosphodiesterase (PDE) domain and an oligonucleotide/oligosaccharide binding (OB) domain. The N-terminal domain and the linker connecting it to the PDE domain are disordered in the reported crystal structure. The p180C adopts an elongated asymmetric saddle shape, with a three-helix bundle in the middle and zinc-binding modules (Zn1 and Zn2) on each side. The extensive p180C-p70 interactions involve 20 hydrogen bonds and a number of hydrophobic interactions resulting in an extended buried surface of 4080 Å(2). Importantly, in the structure of the p180C-p70 complex with full-length p70, the residues from the N-terminal to the OB domain contribute to interactions with p180C. The comparative structural analysis revealed both the conserved features and the differences between the human and yeast Pol α complexes.
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Suwa
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and.
authors.forename: Jianyou
authors.initials: J
authors.lastname: Gu
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and.
authors.forename: Andrey G
authors.initials: AG
authors.lastname: Baranovskiy
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and.
authors.forename: Nigar D
authors.initials: ND
authors.lastname: Babayeva
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and the Departments of Biochemistry and Molecular Biology and Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198.
authors.forename: Youri I
authors.initials: YI
authors.lastname: Pavlov
authors.affiliation: From the Eppley Institute for Research in Cancer and Allied Diseases and ttahirov@unmc.edu.
authors.forename: Tahir H
authors.initials: TH
authors.lastname: Tahirov
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004256:DNA Polymerase I / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000737:chemistry; D016415:Sequence Alignment
chemical_list: D021122:Protein Subunits; D004256:DNA Polymerase I
keywords: DNA polymerase; DNA replication; crystal structure; human genetics; protein complex; zinc finger
references.citation: J Biol Chem. 1999 Aug 6;274(32):22283-8
references.pmid: 10428796
references.citation: Mol Cell Biol. 1999 Nov;19(11):7886-96
references.pmid: 10523676
references.citation: Trends Biochem Sci. 2000 Nov;25(11):572-6
references.pmid: 11084371
references.citation: Nucleic Acids Res. 2004 Jan 22;32(2):e19
references.pmid: 14739232
references.citation: Genes Dev. 2004 May 1;18(9):992-1006
references.pmid: 15132993
references.citation: Nucleic Acids Res. 2004 Jun 01;32(10):3005-16
references.pmid: 15173383
references.citation: Mol Cell Biol. 2004 Sep;24(17):7419-34
references.pmid: 15314153
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Adv Protein Chem. 2004;69:167-203
references.pmid: 15588843
references.citation: Crit Rev Biochem Mol Biol. 2005 Mar-Apr;40(2):115-28
references.pmid: 15814431
references.citation: Curr Biol. 2006 Jan 24;16(2):202-7
references.pmid: 16431373
references.citation: Protein Expr Purif. 2007 May;53(1):40-50
references.pmid: 17251035
references.citation: Nature. 2007 Jul 12;448(7150):157-62
references.pmid: 17581590
references.citation: Proteins. 2008 Feb 15;70(3):611-25
references.pmid: 17876813
references.citation: Cell Res. 2008 Jan;18(1):174-83
references.pmid: 18157155
references.citation: Mol Cell. 2008 Apr 25;30(2):137-44
references.pmid: 18439893
references.citation: Nat Rev Genet. 2008 Aug;9(8):594-604
references.pmid: 18626473
references.citation: Nucleic Acids Res. 2008 Sep;36(15):5102-10
references.pmid: 18676977
references.citation: Cell Cycle. 2008 Oct;7(19):3026-36
references.pmid: 18818516
references.citation: J Biol Chem. 1991 Apr 25;266(12):7893-903
references.pmid: 1902230
references.citation: Biol Direct. 2009 Mar 18;4:11
references.pmid: 19296856
references.citation: EMBO J. 2009 Jul 8;28(13):1978-87
references.pmid: 19494830
references.citation: Mutat Res. 2010 Mar 1;685(1-2):45-53
references.pmid: 19682465
references.citation: BMC Mol Biol. 2009 Aug 17;10:82
references.pmid: 19686603
references.citation: Genetics. 2010 Jan;184(1):27-42
references.pmid: 19841096
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
references.pmid: 20124702
references.citation: Trends Biochem Sci. 2010 Jun;35(6):339-47
references.pmid: 20163964
references.citation: J Biol Chem. 2010 May 28;285(22):17112-22
references.pmid: 20234039
references.citation: Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4601-5
references.pmid: 2052544
references.citation: J Biol Chem. 2010 Oct 22;285(43):33475-84
references.pmid: 20685648
references.citation: Semin Cancer Biol. 2010 Oct;20(5):281-93
references.pmid: 20951805
references.citation: Nat Chem Biol. 2011 Nov 27;8(1):125-32
references.pmid: 22119860
references.citation: J Biol Chem. 2012 May 18;287(21):17281-7
references.pmid: 22465957
references.citation: J Biol Chem. 2012 Jul 6;287(28):23740-7
references.pmid: 22593576
references.citation: J Biol Chem. 2012 Aug 3;287(32):26854-66
references.pmid: 22700977
references.citation: Proc Natl Acad Sci U S A. 2012 Jul 31;109(31):12455-60
references.pmid: 22711820
references.citation: Subcell Biochem. 2012;62:157-69
references.pmid: 22918585
references.citation: Nucleic Acids Res. 2012 Dec;40(22):11618-26
references.pmid: 23066099
references.citation: Curr Biol. 2013 Apr 8;23(7):543-52
references.pmid: 23499531
references.citation: Proc Natl Acad Sci U S A. 2013 Oct 1;110(40):15961-6
references.pmid: 24043831
references.citation: Nucleic Acids Res. 2014 Jan;42(1):290-306
references.pmid: 24049079
references.citation: Proc Natl Acad Sci U S A. 2014 Feb 25;111(8):2954-9
references.pmid: 24449906
references.citation: Curr Opin Genet Dev. 2014 Feb;24:107-13
references.pmid: 24583393
references.citation: J Biol Chem. 2014 Aug 8;289(32):22021-34
references.pmid: 24962573
references.citation: Nat Struct Mol Biol. 2014 Aug;21(8):649-51
references.pmid: 24997601
references.citation: Proc Natl Acad Sci U S A. 2014 Oct 28;111(43):15390-5
references.pmid: 25313033
references.citation: Nucleic Acids Res. 2014 Dec 16;42(22):14013-21
references.pmid: 25429975
references.citation: J Biol Chem. 2015 Feb 27;290(9):5635-46
references.pmid: 25550159
references.citation: Nature. 2015 Feb 26;518(7540):502-506
references.pmid: 25624100
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Mol Cell Biol. 1995 Feb;15(2):883-91
references.pmid: 7823954
references.citation: Mol Cell Biol. 1994 Feb;14(2):923-33
references.pmid: 8289832
references.citation: Nucleic Acids Res. 1997 Mar 15;25(6):1094-9
references.pmid: 9092615
references.citation: Genetics. 1997 Aug;146(4):1239-51
references.pmid: 9258670
references.citation: Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
references.pmid: 9757107
references.citation: J Biol Chem. 1998 Nov 6;273(45):30046-55
references.pmid: 9792727