Human DNA polymerase η is pre-aligned for dNTP binding and catalysis

Ajay Ummat, Timothy D. Silverstein, Rinku Jain, Angeliki Buku, Robert E. Johnson, Louise Prakash, Satya Prakash, Aneel K. Aggarwal

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Pre-steady-state kinetic studies on Y-family DNA polymerase η (Polη) have suggested that the polymerase undergoes a rate-limiting conformational change step before the phosphoryl transfer of the incoming nucleotide to the primer terminus. However, the nature of this rate-limiting conformational change step has been unclear, due in part to the lack of structural information on the Polη binary complex. We present here for the first time a crystal structure of human Polη (hPolη) in binary complex with its DNA substrate. We show that the hPolη domains move only slightly on dNTP binding and that the polymerase by and large is pre-aligned for dNTP binding and catalysis. We also show that there is no major reorientation of the DNA from a nonproductive to a productive configuration and that the active site is devoid of metals in the absence of dNTP. Together, these observations lead us to suggest that the rate-limiting conformational change step in the Polη replication cycle likely corresponds to a rate-limiting entry of catalytic metals in the active site.

Original languageEnglish (US)
Pages (from-to)627-634
Number of pages8
JournalJournal of Molecular Biology
Volume415
Issue number4
DOIs
StatePublished - Jan 27 2012

Fingerprint

DNA-Directed DNA Polymerase
Catalysis
Catalytic Domain
Metals
DNA
Nucleotides

Keywords

  • binding and catalysis
  • DNA polymerase
  • DNA repair
  • translesion DNA synthesis
  • Y-family DNA polymerase

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Ummat, A., Silverstein, T. D., Jain, R., Buku, A., Johnson, R. E., Prakash, L., ... Aggarwal, A. K. (2012). Human DNA polymerase η is pre-aligned for dNTP binding and catalysis. Journal of Molecular Biology, 415(4), 627-634. https://doi.org/10.1016/j.jmb.2011.11.038

Human DNA polymerase η is pre-aligned for dNTP binding and catalysis. / Ummat, Ajay; Silverstein, Timothy D.; Jain, Rinku; Buku, Angeliki; Johnson, Robert E.; Prakash, Louise; Prakash, Satya; Aggarwal, Aneel K.

In: Journal of Molecular Biology, Vol. 415, No. 4, 27.01.2012, p. 627-634.

Research output: Contribution to journalArticle

Ummat, A, Silverstein, TD, Jain, R, Buku, A, Johnson, RE, Prakash, L, Prakash, S & Aggarwal, AK 2012, 'Human DNA polymerase η is pre-aligned for dNTP binding and catalysis', Journal of Molecular Biology, vol. 415, no. 4, pp. 627-634. https://doi.org/10.1016/j.jmb.2011.11.038
Ummat, Ajay ; Silverstein, Timothy D. ; Jain, Rinku ; Buku, Angeliki ; Johnson, Robert E. ; Prakash, Louise ; Prakash, Satya ; Aggarwal, Aneel K. / Human DNA polymerase η is pre-aligned for dNTP binding and catalysis. In: Journal of Molecular Biology. 2012 ; Vol. 415, No. 4. pp. 627-634.
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