Yeast DNA polymerase eta makes functional contacts with the DNA minor groove only at the incoming nucleoside triphosphate.
DNA polymerase eta (Pol eta) functions in the proficient bypass of a variety of DNA lesions. Relative to the replicative polymerases, Pol eta has a greater tolerance for distorted DNA geometries and possesses a low fidelity. X-ray crystal structures and studies with nucleotide analogs have implicated interactions with the DNA minor groove as being crucial for the high fidelity of replicative DNA polymerases. To determine whether Pol eta also makes such functionally important contacts with the DNA minor groove, here we examine the effects on Pol eta-catalyzed nucleotide incorporation when 3-deazaguanine, a base analog that lacks the ability to form minor-groove hydrogen bonds with the protein, is substituted for guanine at various positions in the DNA. From these studies, we conclude that Pol eta makes only a single functional contact with the DNA minor groove at the position of the incoming nucleotide; in this regard, Pol eta differs from high-fidelity DNA polymerases that are unable to replicate through DNA lesions. These results help explain the proficient ability of Pol eta for bypassing distorting DNA lesions.
Washington
M Todd
MT
Sealy Center for Molecular Science, University of Texas Medical Branch, 6.104 Blocker Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555-1061, USA.
Wolfle
William T
WT
Spratt
Thomas E
TE
Prakash
Louise
L
Prakash
Satya
S
D001483:Base Sequence; D017931:DNA Primers; D004271:DNA, Fungal / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D009705:Nucleosides / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D000099270:Y-Family DNA Polymerases
D017931:DNA Primers; D004271:DNA, Fungal; D004259:DNA-Directed DNA Polymerase; D009705:Nucleosides; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases