Primer initiation and extension by T7 DNA primase.

The EMBO Journal
Udi QimronCharles C Richardson

Abstract

T7 DNA primase is composed of a catalytic RNA polymerase domain (RPD) and a zinc-binding domain (ZBD) connected by an unstructured linker. The two domains are required to initiate the synthesis of the diribonucleotide pppAC and its extension into a functional primer pppACCC (de novo synthesis), as well as for the extension of exogenous AC diribonucleotides into an ACCC primer (extension synthesis). To explore the mechanism underlying the RPD and ZBD interactions, we have changed the length of the linker between them. Wild-type T7 DNA primase is 10-fold superior in de novo synthesis compared to T7 DNA primase having a shorter linker. However, the primase having the shorter linker exhibits a two-fold enhancement in its extension synthesis. T7 DNA primase does not catalyze extension synthesis by a ZBD of one subunit acting on a RPD of an adjacent subunit (trans mode), whereas de novo synthesis is feasible in this mode. We propose a mechanism for primer initiation and extension based on these findings.

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Citations

Jan 15, 2008·Nature Structural & Molecular Biology·Jacob E CornJames M Berger
May 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Sebastian GeibelWolfram Saenger
May 31, 2012·Proceedings of the National Academy of Sciences of the United States of America·Arkadiusz W Kulczyk, Charles C Richardson
May 19, 2012·The Journal of Biological Chemistry·Huidong ZhangCharles C Richardson
Oct 2, 2012·The Journal of Biological Chemistry·Seung-Joo LeeCharles C Richardson
Apr 13, 2010·The Journal of Biological Chemistry·Samir M Hamdan, Antoine M van Oijen
Mar 31, 2010·Nucleic Acids Research·Seung-Joo LeeCharles C Richardson
Mar 21, 2009·Annual Review of Biochemistry·Samir M Hamdan, Charles C Richardson
May 14, 2010·Annual Review of Biophysics·Antoine M van Oijen, Joseph J Loparo
Sep 13, 2011·Current Opinion in Chemical Biology·Seung-Joo Lee, Charles C Richardson
Jun 23, 2009·Biochimica Et Biophysica Acta·Robert D Kuchta, Gudrun Stengel
Jun 4, 2015·Annual Review of Biochemistry·Charles C Richardson
May 11, 2016·Proceedings of the National Academy of Sciences of the United States of America·Alfredo J HernandezCharles C Richardson
Aug 14, 2012·The Journal of Biological Chemistry·Huidong ZhangCharles C Richardson
Aug 8, 2014·Molecular Biology Reports·Jamie B Towle-WeickselDonald J Crampton

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