Investigation of mycobacterial <i>recA</i> function: protein introns in the RecA of pathogenic mycobacteria do not affect competency for homologous recombination

Frischkorn, Klaus; Sander, Peter; Scholz, Matthias; Teschner, K; Prammananan, Therdsak; Böttger, Erik C.

doi:10.1046/j.1365-2958.1998.01003.x

Cited by 37 publications

(32 citation statements)

References 43 publications

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“…Instead, M. canettii A was found to be able to successfully transfer DNA into the recipient M. canettii L despite predictions that M. canettii A-derived sequences only made up a minor part of other M. canettii genomes in in silico analyses (11). The impact of inteins on recombination in selected mycobacterial strains might thus be limited, in accordance with previous findings (40).…”

Section: Discussionsupporting

confidence: 78%

Key experimental evidence of chromosomal DNA transfer among selected tuberculosis-causing mycobacteria

Boritsch

Khanna

Pawlik

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

101

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Horizontal gene transfer (HGT) is a major driving force of bacterial diversification and evolution. For tuberculosis-causing mycobacteria, the impact of HGT in the emergence and distribution of dominant lineages remains a matter of debate. Here, by using fluorescenceassisted mating assays and whole genome sequencing, we present unique experimental evidence of chromosomal DNA transfer between tubercle bacilli of the early-branching Mycobacterium canettii clade. We found that the obtained recombinants had received multiple donor-derived DNA fragments in the size range of 100 bp to 118 kbp, fragments large enough to contain whole operons. Although the transfer frequency between M. canettii strains was low and no transfer could be observed among classical Mycobacterium tuberculosis complex (MTBC) strains, our study provides the proof of concept for genetic exchange in tubercle bacilli. This outstanding, now experimentally validated phenomenon presumably played a key role in the early evolution of the MTBC toward pathogenicity. Moreover, our findings also provide important information for the risk evaluation of potential transfer of drug resistance and fitness mutations among clinically relevant mycobacterial strains.recombination | DNA transfer | tuberculosis | Mycobacterium canettii | evolution

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Section: Discussionsupporting

confidence: 78%

Key experimental evidence of chromosomal DNA transfer among selected tuberculosis-causing mycobacteria

Boritsch

Khanna

Pawlik

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

101

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“…1). These results indicated that the recA gene is present in all 36 of the species, a result probably related to the essentiality of RecA recombinase in mycobacteria, as suggested by Frischkorn and collaborators and Papavinasasundaram and collaborators (14,29). Moreover, the amplification yields were rather high, suggesting that the sensitivity of the PCR diagnosis would be quite good, even if the target sequences are present in single copy in the M. tuberculosis genome.…”

Section: Discussionsupporting

confidence: 52%

Specificities and Functions of the recA and pps1 Intein Genes of Mycobacterium tuberculosis and Application for Diagnosis of Tuberculosis

et al. 2002

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The worldwide recrudescence of tuberculosis and the widespread appearance of antibiotic resistance have strengthened the need for rapid and specific diagnostic tools. The prevailing microbiological identification of Mycobacterium tuberculosis, the causative agent of tuberculosis, which implies the use of in vitro cultures and acid-fast staining microscopy, is time-consuming. Detection of M. tuberculosis directly in clinical samples through PCR amplification of mycobacterium-specific genes, designed to shorten diagnostic delay, demonstrated reliability and high sensitivity. However, the quality of the diagnosis depends on the specificity of the target sequence for M. tuberculosis complex strains. In the present study, we demonstrated the specificity of recA and pps1 inteins for this complex and thus the feasibility of using intein-coding sequences as a new target for PCR diagnosis. Indeed, the recA and pps1 genes of 36 clinical isolates of M. tuberculosis and 10 field strains of M. bovis were found to be interrupted by an intein sequence at the RecA-a and Pps1-b sites, respectively, while a large number of nontuberculous mycobacterial species failed to demonstrate these insertions. Besides, the MtuPps1, which was cloned and expressed in Escherichia coli, was shown to possess an endonuclease activity. The intein cleaves the 40-bp sequence spanning the intein insertion site Pps1-b in the inteinless pps1 gene. In addition to the PCR amplification of recA and pps1 intein genes as a tool for diagnosis, the specific endonuclease activity could represent a new molecular approach to identify M. tuberculosis.Tuberculosis remains a major public health problem in developing countries. Despite the decline in incidence observed in the 1980s, resurgence has recently occurred. Furthermore, the recrudescence of infections with Mycobacterium tuberculosis and the emergence of multidrug-resistant strains represent a serious public health threat in developed countries, where the incidence of tuberculosis is related to immunodeficient patients, homeless people, and the advancing age of the population (World Health Organization web site at http://www .who.int/health-topics/tb.htm).This worldwide resurgence of tuberculosis has strengthened the need for rapid and specific diagnostic tools to control its spread. Delay in diagnosis results, first, in late initiation of antitubercular therapy and, second, in prolonged transmission of the infection.Until recently, the diagnosis of tuberculosis was based on clinical features and microbiological assays, radiological examinations, immunological tests, microscopy identification, or in vitro cultures (37). Acid-fast staining microscopy of specimens combined with isolation and culture of the bacilli remains the "gold standard" method to specifically identify mycobacteria. Because of the low growth rate of M. tuberculosis, this method is time-consuming, and the diagnosis can take up to 8 weeks.More recently, molecular approaches have been designed to ensure early detection of M. tuberculosis in cl...

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“…They can be found in proteins involved in DNA or RNA metabolism and biosynthesis (such as a ribonucleotide reductase in Anabaena [139]); cell division (such as an ATPase involved in chromatin remodeling in Deinococcus radiodurans [140]); transcription (such as the GyrA gyrase in mycobacteria [141]); and DNA replication (such as DnaX, the DNA polymerase subunit, in Synechocystis and the DnaB helicase in Rhodothermus marinus [142,143]), repair, and recombination (such as RecA in mycobacteria [144]). There are several nonexclusive hypotheses for their localization.…”

Section: Inteins Introns and Retroelementsmentioning

confidence: 99%

Bacterial Genome Instability

Darmon

Leach

2014

Microbiol Mol Biol Rev

348

289

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SUMMARY Bacterial genomes are remarkably stable from one generation to the next but are plastic on an evolutionary time scale, substantially shaped by horizontal gene transfer, genome rearrangement, and the activities of mobile DNA elements. This implies the existence of a delicate balance between the maintenance of genome stability and the tolerance of genome instability. In this review, we describe the specialized genetic elements and the endogenous processes that contribute to genome instability. We then discuss the consequences of genome instability at the physiological level, where cells have harnessed instability to mediate phase and antigenic variation, and at the evolutionary level, where horizontal gene transfer has played an important role. Indeed, this ability to share DNA sequences has played a major part in the evolution of life on Earth. The evolutionary plasticity of bacterial genomes, coupled with the vast numbers of bacteria on the planet, substantially limits our ability to control disease.

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Investigation of mycobacterial recA function: protein introns in the RecA of pathogenic mycobacteria do not affect competency for homologous recombination

Cited by 37 publications

References 43 publications

Key experimental evidence of chromosomal DNA transfer among selected tuberculosis-causing mycobacteria

Key experimental evidence of chromosomal DNA transfer among selected tuberculosis-causing mycobacteria

Specificities and Functions of the recA and pps1 Intein Genes of Mycobacterium tuberculosis and Application for Diagnosis of Tuberculosis

Bacterial Genome Instability

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