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Title: Crosstalk between primase subunits can act to regulate primersynthesis in trans

Abstract

The coordination of primase function within the replisome is an essential but poorly understood feature of lagging strand synthesis. By using crystallography and small-angle X-ray scattering (SAXS), we show that functional elements of bacterial primase transition between two dominant conformations: an extended form that uncouples a regulatory domain from its associated RNA polymerase core and a compact state that sequesters the regulatory region from the site of primer synthesis. FRET studies and priming assays reveal that the regulatory domain of one primase subunit productively associates with nucleic acid that is bound to the polymerase domain of a second protomer in trans. This intersubunit interaction allows primase to select initiation sites on template DNA and implicates the regulatory domain as a 'molecular brake' that restricts primer length. Our data suggest that the replisome may cooperatively use multiple primases and this conformational switch to control initiation frequency, processivity, and ultimately, Okazaki fragment synthesis.

Authors:
; ; ;
Publication Date:
Research Org.:
Ernest Orlando Lawrence Berkeley NationalLaboratory, Berkeley, CA (US)
Sponsoring Org.:
USDOE Director. Office of Science. Biological andEnvironmental Research
OSTI Identifier:
918923
Report Number(s):
LBNL-61529
R&D Project: 441H01; BnR: KP1101010; TRN: US200819%%500
DOE Contract Number:  
DE-AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
Molecular Cell
Additional Journal Information:
Journal Volume: 20; Related Information: Journal Publication Date: November 11,2005
Country of Publication:
United States
Language:
English
Subject:
59; CRYSTALLOGRAPHY; DNA; FUNCTIONALS; NUCLEIC ACIDS; POLYMERASES; RNA POLYMERASES; SCATTERING; SYNTHESIS

Citation Formats

Corn, Jacob E, Pease, Paul J, Hura, Greg L, and Berger, James M. Crosstalk between primase subunits can act to regulate primersynthesis in trans. United States: N. p., 2005. Web. doi:10.1016/j.molcel.2005.09.004.
Corn, Jacob E, Pease, Paul J, Hura, Greg L, & Berger, James M. Crosstalk between primase subunits can act to regulate primersynthesis in trans. United States. doi:10.1016/j.molcel.2005.09.004.
Corn, Jacob E, Pease, Paul J, Hura, Greg L, and Berger, James M. Fri . "Crosstalk between primase subunits can act to regulate primersynthesis in trans". United States. doi:10.1016/j.molcel.2005.09.004.
@article{osti_918923,
title = {Crosstalk between primase subunits can act to regulate primersynthesis in trans},
author = {Corn, Jacob E and Pease, Paul J and Hura, Greg L and Berger, James M},
abstractNote = {The coordination of primase function within the replisome is an essential but poorly understood feature of lagging strand synthesis. By using crystallography and small-angle X-ray scattering (SAXS), we show that functional elements of bacterial primase transition between two dominant conformations: an extended form that uncouples a regulatory domain from its associated RNA polymerase core and a compact state that sequesters the regulatory region from the site of primer synthesis. FRET studies and priming assays reveal that the regulatory domain of one primase subunit productively associates with nucleic acid that is bound to the polymerase domain of a second protomer in trans. This intersubunit interaction allows primase to select initiation sites on template DNA and implicates the regulatory domain as a 'molecular brake' that restricts primer length. Our data suggest that the replisome may cooperatively use multiple primases and this conformational switch to control initiation frequency, processivity, and ultimately, Okazaki fragment synthesis.},
doi = {10.1016/j.molcel.2005.09.004},
journal = {Molecular Cell},
number = ,
volume = 20,
place = {United States},
year = {2005},
month = {8}
}