Cloning and characterization of the Haemophilus influenzae mutB gene

Stuy, Johan H.; Walter, Ronald B.

doi:10.1128/jb.175.16.5265-5267.1993

Cited by 1 publication

(1 citation statement)

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“…In another study, inactivation of Haemophilus influenzae uvrD led to a two‐fold increase in spontaneous mutation resulting in rifampicin‐resistance and a three‐fold increase in rates of pilus phase variation due to the destabilization of long dinucleotide repeat tracts [27]. Furthermore, Haemophilus influenzae uvrD could complement E. coli uvrD mutants [28]. Hence, UvrD constitutes an attractive drug target for the development of new antibacterial agents to combat these pathogens.…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of UvrD helicases from Haemophilus influenzae and Helicobacter pylori

Sharma

Rao

2012

The FEBS Journal

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Haemophilus influenzae and Helicobacter pylori are major bacterial pathogens that face high levels of genotoxic stress within their host. UvrD, a ubiquitous bacterial helicase that plays important roles in multiple DNA metabolic pathways, is essential for genome stability and might, therefore, be crucial in bacterial physiology and pathogenesis. In this study, the functional characterization of UvrD helicase from Haemophilus influenzae and Helicobacter pylori is reported. UvrD from Haemophilus influenzae (HiUvrD) and Helicobacter pylori (HpUvrD) exhibit strong single‐stranded DNA‐specific ATPase and 3′–5′ helicase activities. Mutation of highly conserved arginine (R288) in HiUvrD and glutamate (E206) in HpUvrD abrogated their activities. Both the proteins were able to bind and unwind a variety of DNA structures including duplexes with strand discontinuities and branches, three‐ and four‐way junctions that underpin their role in DNA replication, repair and recombination. HiUvrD required a minimum of 12 nucleotides, whereas HpUvrD preferred 20 or more nucleotides of 3′‐single‐stranded DNA tail for efficient unwinding of duplex DNA. Interestingly, HpUvrD was able to hydrolyze and utilize GTP for its helicase activity although not as effectively as ATP, which has not been reported to date for UvrD characterized from other organisms. HiUvrD and HpUvrD were found to exist predominantly as monomers in solution together with multimeric forms. Noticeably, deletion of distal C‐terminal 48 amino acid residues disrupted the oligomerization of HiUvrD, whereas deletion of 63 amino acids from C‐terminus of HpUvrD had no effect on its oligomerization. This study presents the characteristic features and comparative analysis of Haemophilus influenzae and Helicobacter pylori UvrD, and constitutes the basis for understanding the role of UvrD in the biology and virulence of these pathogens. Structured digital abstract http://www.uniprot.org/uniprot/Q02322 and http://www.uniprot.org/uniprot/Q02322 http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0407 by http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0071 (http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-8361685) http://www.uniprot.org/uniprot/O26013 and http://www.uniprot.org/uniprot/O26013 http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0407 by http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0071 (http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-8361776) http://www.uniprot.org/uniprot/Q02322 and http://www.uniprot.org/uniprot/Q02322 http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0407 by http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0030 (http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-8361717) http://www.uniprot.org/uniprot/O26013 and http://www.uniprot.org/uniprot/O26013 http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0407 by http://www.ebi.ac.uk/ontology-lookup/?termId=MI:0030 (http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-8361732)

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Section: Introductionmentioning

confidence: 99%