Mechanism of DNA Polymerization Catalyzed bySulfolobus solfataricusP2 DNA Polymerase IV†

Biochemistry ◽  
2004 ◽  
Vol 43 (7) ◽  
pp. 2116-2125 ◽  
Author(s):  
Kevin A. Fiala ◽  
Zucai Suo
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerization ◽  
Dna Polymerase Iv

scholarly journals Interactions and Localization of Escherichia coli Error-Prone DNA Polymerase IV after DNA Damage

2015 ◽  
Vol 197 (17) ◽  
pp. 2792-2809 ◽  
Author(s):  
Sarita Mallik ◽  
Ellen M. Popodi ◽  
Andrew J. Hanson ◽  
Patricia L. Foster
Keyword(s):  
Dna Damage ◽  
Dna Polymerase ◽  
Dna Helicase ◽  
Dna Lesions ◽  
Replication Forks ◽  
Content Type ◽  
Pol Iv ◽  
Dna Polymerase Iv ◽  
Stalled Replication Forks

ABSTRACTEscherichia coli's DNA polymerase IV (Pol IV/DinB), a member of the Y family of error-prone polymerases, is induced during the SOS response to DNA damage and is responsible for translesion bypass and adaptive (stress-induced) mutation. In this study, the localization of Pol IV after DNA damage was followed using fluorescent fusions. After exposure ofE. colito DNA-damaging agents, fluorescently tagged Pol IV localized to the nucleoid as foci. Stepwise photobleaching indicated ∼60% of the foci consisted of three Pol IV molecules, while ∼40% consisted of six Pol IV molecules. Fluorescently tagged Rep, a replication accessory DNA helicase, was recruited to the Pol IV foci after DNA damage, suggesting that thein vitrointeraction between Rep and Pol IV reported previously also occursin vivo. Fluorescently tagged RecA also formed foci after DNA damage, and Pol IV localized to them. To investigate if Pol IV localizes to double-strand breaks (DSBs), an I-SceI endonuclease-mediated DSB was introduced close to a fluorescently labeled LacO array on the chromosome. After DSB induction, Pol IV localized to the DSB site in ∼70% of SOS-induced cells. RecA also formed foci at the DSB sites, and Pol IV localized to the RecA foci. These results suggest that Pol IV interacts with RecAin vivoand is recruited to sites of DSBs to aid in the restoration of DNA replication.IMPORTANCEDNA polymerase IV (Pol IV/DinB) is an error-prone DNA polymerase capable of bypassing DNA lesions and aiding in the restart of stalled replication forks. In this work, we demonstratein vivolocalization of fluorescently tagged Pol IV to the nucleoid after DNA damage and to DNA double-strand breaks. We show colocalization of Pol IV with two proteins: Rep DNA helicase, which participates in replication, and RecA, which catalyzes recombinational repair of stalled replication forks. Time course experiments suggest that Pol IV recruits Rep and that RecA recruits Pol IV. These findings providein vivoevidence that Pol IV aids in maintaining genomic stability not only by bypassing DNA lesions but also by participating in the restoration of stalled replication forks.

scholarly journals SOS Mutator DNA Polymerase IV Functions in Adaptive Mutation and Not Adaptive Amplification

2001 ◽  
Vol 7 (3) ◽  
pp. 571-579 ◽  
Author(s):  
Gregory J McKenzie ◽  
Peter L Lee ◽  
Mary-Jane Lombardo ◽  
P.J Hastings ◽  
Susan M Rosenberg
Keyword(s):  
Dna Polymerase ◽  
Adaptive Mutation ◽  
Dna Polymerase Iv

Synthesis of N2-Deoxyguanosine Modified DNAs and the Studies on Their Translesion Synthesis by the E. coli DNA Polymerase IV

2019 ◽  
Vol 84 (4) ◽  
pp. 1734-1747 ◽  
Author(s):  
Pratibha P. Ghodke ◽  
Praneeth Bommisetti ◽  
Deepak T. Nair ◽  
P. I. Pradeepkumar
Keyword(s):  
Dna Polymerase ◽  
Translesion Synthesis ◽  
E Coli ◽  
Dna Polymerase Iv

scholarly journals Single-molecule live-cell imaging reveals RecB-dependent function of DNA polymerase IV in double strand break repair

2020 ◽  
Vol 48 (15) ◽  
pp. 8490-8508 ◽  
Author(s):  
Sarah S Henrikus ◽  
Camille Henry ◽  
Amy E McGrath ◽  
Slobodan Jergic ◽  
John P McDonald ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Single Molecule ◽  
Strand Break ◽  
Double Strand Break ◽  
Double Strand Breaks ◽  
Strand Breaks ◽  
E Coli ◽  
Pol Iv ◽  
Dna Polymerase Iv

Abstract Several functions have been proposed for the Escherichia coli DNA polymerase IV (pol IV). Although much research has focused on a potential role for pol IV in assisting pol III replisomes in the bypass of lesions, pol IV is rarely found at the replication fork in vivo. Pol IV is expressed at increased levels in E. coli cells exposed to exogenous DNA damaging agents, including many commonly used antibiotics. Here we present live-cell single-molecule microscopy measurements indicating that double-strand breaks induced by antibiotics strongly stimulate pol IV activity. Exposure to the antibiotics ciprofloxacin and trimethoprim leads to the formation of double strand breaks in E. coli cells. RecA and pol IV foci increase after treatment and exhibit strong colocalization. The induction of the SOS response, the appearance of RecA foci, the appearance of pol IV foci and RecA-pol IV colocalization are all dependent on RecB function. The positioning of pol IV foci likely reflects a physical interaction with the RecA* nucleoprotein filaments that has been detected previously in vitro. Our observations provide an in vivo substantiation of a direct role for pol IV in double strand break repair in cells treated with double strand break-inducing antibiotics.

scholarly journals Optimal numbers of residues in linkers of DNA polymerase I, T7 primase and DNA polymerase IV

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Yi-Ben Fu ◽  
Zhan-Feng Wang ◽  
Peng-Ye Wang ◽  
Ping Xie
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase I ◽  
Dna Polymerase Iv ◽  
Polymerase I

scholarly journals Comparison of the in Vitro Replication of the 7-(2-Oxoheptyl)-1,N2-etheno-2′-deoxyguanosine and 1,N2-Etheno-2′-deoxyguanosine Lesions bySulfolobus solfataricusP2 DNA Polymerase IV (Dpo4)

2010 ◽  
Vol 23 (8) ◽  
pp. 1330-1341 ◽  
Author(s):  
Plamen P. Christov ◽  
Katya V. Petrova ◽  
Ganesh Shanmugam ◽  
Ivan D. Kozekov ◽  
Albena Kozekova ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
In Vitro Replication ◽  
Dna Polymerase Iv
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