Processing of A-form ssDNA by cryptic RNase H fold exonuclease PF2046

Abstract

RNase H fold protein PF2046 of Pyrococcus furiosus is a 3'-5' ssDNA exonuclease that cleaves after the second nucleotide from the 3' end of ssDNA and prefers poly-dT over poly-dA as a substrate. In our crystal structure of PF2046 complexed with an oligonucleotide of four thymidine nucleotides (dT(4)), PF2046 accommodates dT(4) tightly in a groove and imposes steric hindrance on dT(4) mainly by Phe220 such that dT(4) assumes the A-form. As poly-dA prefer B-form due to the stereochemical restrictions, the A-form ssDNA binding by PF2046 should disfavor the processing of poly-dA. Phe220 variants display reduced activity toward poly-dA and the A-form appears to be a prerequisite for the processing by PF2046. (C) 2016 Elsevier Inc. All rights reserved.