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Role of Pol κ and DinB distal residues on extension step of TLS
Author(s) -
Stern Hannah R,
Mills Caitlyn L,
Ondrechen Mary Jo,
Beuning Penny J
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.592.6
Subject(s) - dna polymerase , polymerase , biology , dna damage , dna , dna polymerase ii , genetics , polymerase chain reaction , reverse transcriptase , gene
DNA is subject to damage from endogenous and exogenous sources. Replicative DNA polymerases are typically unable to replicate damaged DNA, but specialized DNA polymerases in the Y family possess this ability. Escherichia coli has two Y family polymerases that are specialized to bypass lesions when copying damaged DNA in a process called translesion synthesis. DinB is one of these polymerases, which is involved in bypassing deoxyguanosine adducts at the N2 position. There are also four human Y family polymerases, including DNA polymerase Kappa, that have similar function. However, Pol κ is more efficient in copying past DNA damage lesions in the extension step of translesion synthesis. In order to investigate the importance of particular residues in the extension step of TLS, the computational method POOL was utilized. This tool identified active site residues and residues previously observed to be important for activity. POOL also predicted more distant residues that do not have direct contact with substrate that may have catalytic importance, but the residues are in different regions of DinB and Pol κ. In addition, previous HXMS work done in our lab suggested a possible allosteric change in both DinB and Pol κ in different distal residues that POOL did not predict. To study the contribution of these distal residues on the extension step of TLS, DinB and Pol κ variants with mutations at the predicted distal positions were constructed and are being assayed for bypass of damage lesions. Support or Funding Information Support from NSF‐MCB‐1517290, American Cancer Society RSG‐12‐161‐01‐DMC, and the PhRMA Foundation (predoctoral fellowship in informatics awarded to CLM)