Full metadata record
DC Field | Value | Language |
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dc.contributor.author | Choi, Jeong-Yun | - |
dc.contributor.author | Patra, Amritaj | - |
dc.contributor.author | Yeom, Mina | - |
dc.contributor.author | Lee, Young-Sam | - |
dc.contributor.author | Zhang, Qianqian | - |
dc.contributor.author | Egli, Martin | - |
dc.contributor.author | Guengerich, F. Peter | - |
dc.date.available | 2017-05-11T01:40:54Z | - |
dc.date.created | 2017-04-10 | - |
dc.date.issued | 2016-09 | - |
dc.identifier.issn | 0021-9258 | - |
dc.identifier.uri | http://hdl.handle.net/20.500.11750/1633 | - |
dc.description.abstract | DNA polymerase (pol) ι is a Y-family polymerase involved in translesion synthesis, exhibiting higher catalytic activity with Mn2+ than Mg2+. The human germline R96G variant impairs both Mn2+-dependent and Mg2+-dependent activities of pol ι, whereas the Δ1-25 variant selectively enhances its Mg2+-dependent activity. We analyzed pre-steady-state kinetic and structural effects of these two metal ions and genetic variations on pol ι using pol ι core (residues 1-445) proteins. The presence of Mn2+ (0.15 mM) instead of Mg2+ (2 mM) caused a 770-fold increase in efficiency (kpol/Kd,dCTP) of pol ι for dCTP insertion opposite G, mainly due to a 450-fold decrease in Kd,dCTP. The R96G and Δ1-25 variants displayed a 53-fold decrease and a 3-fold increase, respectively, in kpol/Kd,dCTP for dCTP insertion opposite G with Mg2+ when compared with wild type, substantially attenuated by substitution with Mn2+. Crystal structures of pol ι ternary complexes, including the primer terminus 3′-OH and a non-hydrolyzable dCTP analogue opposite G with the active-site Mg2+ or Mn2+, revealed that Mn2+ achieves more optimal octahedral coordination geometry than Mg2+, with lower values in average coordination distance geometry in the catalytic metal A-site. Crystal structures of R96G revealed the loss of three H-bonds of residues Gly-96 and Tyr-93 with an incoming dNTP, due to the lack of an arginine, as well as a destabilized Tyr-93 side chain secondary to the loss of a cation-π interaction between both side chains. These results provide a mechanistic basis for alteration in pol ι catalytic function with coordinating metals and genetic variation. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. | - |
dc.publisher | American Society for Biochemistry and Molecular Biology Inc. | - |
dc.title | Kinetic and Structural Impact of Metal Ions and Genetic Variations on Human DNA Polymerase iota | - |
dc.type | Article | - |
dc.identifier.doi | 10.1074/jbc.M116.748285 | - |
dc.identifier.scopusid | 2-s2.0-84988930359 | - |
dc.identifier.bibliographicCitation | Journal of Biological Chemistry, v.291, no.40, pp.21063 - + | - |
dc.description.isOpenAccess | FALSE | - |
dc.subject.keywordPlus | ACTIVE-SITE | - |
dc.subject.keywordPlus | ADDUCTS | - |
dc.subject.keywordPlus | Amino ACIDs | - |
dc.subject.keywordPlus | B-FAMILY | - |
dc.subject.keywordPlus | Catalysis | - |
dc.subject.keywordPlus | Catalyst Activity | - |
dc.subject.keywordPlus | Catalytic Functions | - |
dc.subject.keywordPlus | Cation-Pi Interactions | - |
dc.subject.keywordPlus | Chains | - |
dc.subject.keywordPlus | Coordination Reactions | - |
dc.subject.keywordPlus | Distance Geometry | - |
dc.subject.keywordPlus | ERROR-PRONE BYPASS | - |
dc.subject.keywordPlus | ETA | - |
dc.subject.keywordPlus | Genes | - |
dc.subject.keywordPlus | HUMAN-CELLS | - |
dc.subject.keywordPlus | Human DNA Polymerase | - |
dc.subject.keywordPlus | LESION-BYPASS | - |
dc.subject.keywordPlus | Manganese | - |
dc.subject.keywordPlus | Metal Ions | - |
dc.subject.keywordPlus | METALS | - |
dc.subject.keywordPlus | Octahedral Coordination Geometry | - |
dc.subject.keywordPlus | OPPOSITE | - |
dc.subject.keywordPlus | Polymers | - |
dc.subject.keywordPlus | REPLICATION | - |
dc.subject.keywordPlus | Structural Effect | - |
dc.subject.keywordPlus | Structural Impact | - |
dc.subject.keywordPlus | Translesion Synthesis | - |
dc.citation.endPage | + | - |
dc.citation.number | 40 | - |
dc.citation.startPage | 21063 | - |
dc.citation.title | Journal of Biological Chemistry | - |
dc.citation.volume | 291 | - |