Online citations, reference lists, and bibliographies.
← Back to Search

Reconstitution Of F Factor DNA Replication In Vitro With Purified Proteins*

S. Zzaman, M. Abhyankar, D. Bastia
Published 2004 · Medicine, Biology

Cite This
Download PDF
Analyze on Scholarcy
Share
Jacob, Brenner, and Cuzin pioneered the development of the F plasmid as a model system to study replication control, and these investigations led to the development of the “replicon model” (Jacob, F., Brenner, S., and Cuzin, F. (1964) Cold Spring Harbor Symp. Quant. Biol. 28, 329–348). To elucidate further the mechanism of initiation of replication of this plasmid and its control, we have reconstituted its replication in vitro with 21 purified host-encoded proteins and the plasmid-encoded initiator RepE. The replication in vitro was specifically initiated at the F ori (oriV) and required both the bacterial initiator protein DnaA and the plasmid-encoded initiator RepE. The wild type dimeric RepE was inactive in catalyzing replication, whereas a monomeric mutant form called RepE* (R118P) was capable of catalyzing vigorous replication. The replication topology was mostly of the Cairns form, and the fork movement was unidirectional and mostly from right to left. The replication was dependent on the HU protein, and the structurally and functionally related DNA bending protein IHF could not efficiently substitute for HU. The priming was dependent on DnaG primase. Many of the characteristics of the in vitro replication closely mimicked those of in vivo replication. We believe that the in vitro system should be very useful in unraveling the mechanism of replication initiation and its control.
This paper references
10.1016/S0300-9084(00)01151-2
Overproduction and improved strategies to purify the threenative forms of nuclease-free HU protein.
O. Pellegrini (2000)
10.1074/jbc.272.32.20173
Role of TrfA and DnaA Proteins in Origin Opening during Initiation of DNA Replication of the Broad Host Range Plasmid RK2*
I. Konieczny (1997)
10.1073/PNAS.96.1.73
Mechanism of recruitment of DnaB helicase to the replication origin of the plasmid pSC101.
H. J. Datta (1999)
10.1016/0092-8674(88)90468-0
Mini-P1 plasmid replication: The autoregulation-sequestration paradox
D. Chattoraj (1988)
10.1016/0076-6879(95)62041-9
DNA synthesis initiated at oriC: in vitro replication reactions.
E. Crooke (1995)
10.1093/emboj/18.17.4597
Crystal structure of a prokaryotic replication initiator protein bound to DNA at 2.6 Å resolution
H. Komori (1999)
10.1093/emboj/20.16.4577
Mechanistic aspects of DnaA–RepA interaction as revealed by yeast forward and reverse two‐hybrid analysis
R. Sharma (2001)
Activation of DnaA5 protein by GrpE and DnaK heat shock proteins in initiation of DNA replication in Escherichia coli.
T. R. Hupp (1993)
10.1101/GAD.5.8.1453
Cooperativity at a distance promoted by the combined action of two replication initiator proteins and a DNA bending protein at the replication origin of pSC101.
T. T. Stenzel (1991)
10.1073/PNAS.91.9.3839
Replication initiator protein RepE of mini-F plasmid: functional differentiation between monomers (initiator) and dimers (autogenous repressor).
M. Ishiai (1994)
10.1093/emboj/17.17.5192
Mechanistic studies of initiator–initiator interaction and replication initiation
Y. Lu (1998)
10.1126/SCIENCE.118.3059.169
Sex in bacteria; genetic studies, 1945-1952.
J. Lederberg (1953)
10.1007/s004380050294
The localized melting of mini-F origin by the combined action of the mini-F initiator protein (RepE) and HU and DnaA of Escherichia coli
Y. Kawasaki (1996)
10.1073/PNAS.86.20.7942
Negative control of plasmid R6K replication: possible role of intermolecular coupling of replication origins.
M. Mceachern (1989)
10.1038/158558A0
Gene Recombination in Escherichia Coli
J. Lederberg (1946)
10.1016/S0092-8674(00)81824-3
Crystal Structure of an IHF-DNA Complex: A Protein-Induced DNA U-Turn
P. Rice (1996)
10.1073/PNAS.93.23.12902
Direct physical interaction between DnaG primase and DnaB helicase of Escherichia coli is necessary for optimal synthesis of primer RNA.
Y. Lu (1996)
10.1007/BF00428895
Replication of mini-F plasmid in vitro promoted by purified E protein
K. Muraiso (2004)
10.1016/S0022-2836(61)80072-7
Genetic regulatory mechanisms in the synthesis of proteins.
F. Jacob (1961)
10.1074/JBC.M312052200
Biochemical Investigations of Control of Replication Initiation of Plasmid R6K*
M. Abhyankar (2004)
10.1016/S1097-2765(00)80205-1
Replisome assembly at oriC, the replication origin of E. coli, reveals an explanation for initiation sites outside an origin.
L. Fang (1999)
10.1101/SQB.1963.028.01.048
On the Regulation of DNA Replication in Bacteria
F. Jacob (1963)
10.1107/S0907444999000578
The high-resolution structure of DNA-binding protein HU from Bacillus stearothermophilus.
S. White (1999)
10.1074/JBC.M308516200
Reconstitution of R6K DNA Replication in Vitro Using 22 Purified Proteins*
M. Abhyankar (2003)
10.1016/0092-8674(81)90095-7
Direct repeats of the F plasmid incC region express F incompatibility
A. Tolun (1981)
10.1128/.61.2.212-238.1997
Stable DNA replication: interplay between DNA replication, homologous recombination, and transcription.
T. Kogoma (1997)
10.1007/BF00260494
Roles of Escherichia coli heat shock proteins DnaK, DnaJ and GrpE in mini-F plasmid replication
Y. Kawasaki (2004)
10.1006/JMBI.1997.1373
The central region of RepE initiator protein of mini-F plasmid plays a crucial role in dimerization required for negative replication control.
F. Matsunaga (1997)
Reconstitution of a nine-protein system that initiates bacteriophage lambda DNA replication.
K. Mensa-Wilmot (1989)
10.1007/BF00328075
Identification of the minimal essential region for the replication origin of miniF plasmid
T. Murotsu (2004)
Replication of plasmid R6K origin gamma in vitro. Dependence on dual initiator proteins and inhibition by transcription.
T. MacAllister (1991)
10.1128/JB.174.17.5597-5603.1992
Mini-F plasmid mutants able to replicate in Escherichia coli deficient in the DnaJ heat shock protein.
M. Ishiai (1992)
10.1016/0092-8674(87)90642-8
The localization of replication origins on ARS plasmids in S. cerevisiae
B. Brewer (1987)
10.1093/emboj/18.13.3856
Regulation of DNA replication by iterons: an interaction between the ori2 and incC regions mediated by RepE‐bound iterons inhibits DNA replication of mini‐F plasmid in Escherichia coli
H. Uga (1999)
10.1016/0092-8674(84)90539-7
Replication initiated at the origin (oriC) of the E. coli chromosome reconstituted with purified enzymes
J. Kaguni (1984)
10.1073/PNAS.81.24.7845
Absence of RNase H allows replication of pBR322 in Escherichia coli mutants lacking DNA polymerase I.
T. Kogoma (1984)
10.1128/JB.169.9.4124-4127.1987
Overproduction of Escherichia coli integration host factor, a protein with nonidentical subunits.
H. Nash (1987)
Primase couples leading- and lagging-strand DNA synthesis from oriC.
H. Hiasa (1994)
10.1016/0092-8674(87)90547-2
The integration host factor of Escherichia coli binds to bent DNA at the origin of replication of the plasmid pSC101
T. T. Stenzel (1987)
10.1016/0092-8674(89)90012-3
The replication terminator protein of E. coli is a DNA sequence-specific contra-helicase
G. S. Khatri (1989)
10.1046/j.1365-2958.2000.02096.x
The interaction domains of the DnaA and DnaB replication proteins of Escherichia coli
H. Seitz (2000)
10.1073/PNAS.74.3.1138
Bidirection replication from a unique origin in a mini-F plasmid.
R. Eichenlaub (1977)



This paper is referenced by
10.1016/j.jmb.2016.04.027
Crowding Modulates the Conformation, Affinity, and Activity of the Components of the Bacterial Disaggregase Machinery.
Garbiñe Celaya (2016)
10.1111/j.1574-6976.2011.00300.x
AT-rich region and repeated sequences - the essential elements of replication origins of bacterial replicons.
M. Rajewska (2012)
10.1128/microbiolspec.PLAS-0026-2014
Iteron Plasmids.
I. Konieczny (2014)
10.3389/fmolb.2016.00039
Replisome Assembly at Bacterial Chromosomes and Iteron Plasmids
K. Węgrzyn (2016)
10.1128/AAC.01540-07
Sequence of Conjugative Plasmid pIP1206 Mediating Resistance to Aminoglycosides by 16S rRNA Methylation and to Hydrophilic Fluoroquinolones by Efflux
B. Périchon (2008)
10.1074/jbc.M112.430595
Structural Insights into the Chaperone Activity of the 40-kDa Heat Shock Protein DnaJ
J. Cuéllar (2013)
10.1099/MIC.0.27773-0
The 120 592 bp IncF plasmid pRSB107 isolated from a sewage-treatment plant encodes nine different antibiotic-resistance determinants, two iron-acquisition systems and other putative virulence-associated functions.
R. Szczepanowski (2005)
10.1042/ETLS20190026
Toward long-lasting artificial cells that better mimic natural living cells
Noël Yeh Martín (2019)
10.1002/bies.201400052
Unique features of DNA replication in mitochondria: A functional and evolutionary perspective
I. Holt (2014)
10.3389/fmolb.2016.00062
Opening the Strands of Replication Origins—Still an Open Question
Jyoti K. Jha (2016)
10.1074/jbc.M109.067348
Replication Initiation at a Distance
Mukesh Saxena (2009)
10.1016/j.plasmid.2014.10.003
Two replication initiators - one mechanism for replication origin opening?
Elzbieta Zabrocka (2014)
10.1074/jbc.M407531200
The DnaK-DnaJ-GrpE Chaperone System Activates Inert Wild Type π Initiator Protein of R6K into a Form Active in Replication Initiation*
S. Zzaman (2004)
10.1109/TCBB.2015.2396057
Fast Prediction of DNA Melting Bubbles Using DNA Thermodynamic Stability
Jan Zrimec (2015)
10.1074/jbc.M110.144642
Role of DnaJ G/F-rich Domain in Conformational Recognition and Binding of Protein Substrates*
Judit Perales-Calvo (2010)
10.1016/j.plasmid.2016.05.002
Proteolysis in plasmid DNA stable maintenance in bacterial cells.
Anna Karłowicz (2016)
10.1128/ecosalplus.ESP-0003-2018
Spread and Persistence of Virulence and Antibiotic Resistance Genes: A Ride on the F Plasmid Conjugation Module.
G. Koraimann (2018)
10.1016/J.MOLCEL.2005.10.037
Oligomeric initiator protein-mediated DNA looping negatively regulates plasmid replication in vitro by preventing origin melting.
S. Zzaman (2005)
10.1534/genetics.107.075861
The Escherichia coli Histone-like Protein HU Has a Role in Stationary Phase Adaptive Mutation
A. B. Williams (2007)
10.1074/jbc.M109.067439
Investigations of π Initiator Protein-mediated Interaction between Replication Origins α and γ of the Plasmid R6K*
Mukesh Saxena (2009)
Semantic Scholar Logo Some data provided by SemanticScholar