Mycobacterium tuberculosis RecG Protein but Not RuvAB or RecA Protein Is Efficient at Remodeling the Stalled Replication Forks IMPLICATIONS FOR MULTIPLE MECHANISMS OF REPLICATION RESTART IN MYCOBACTERIA


Autoria(s): Thakur, Roshan Singh; Basavaraju, Shivakumar; Khanduja, Jasbeer Singh; Muniyappa, K; Nagaraju, Ganesh
Data(s)

2015

Resumo

Aberrant DNA replication, defects in the protection, and restart of stalled replication forks are major causes of genome instability in all organisms. Replication fork reversal is emerging as an evolutionarily conserved physiological response for restart of stalled forks. Escherichia coli RecG, RuvAB, and RecA proteins have been shown to reverse the model replication fork structures in vitro. However, the pathways and the mechanisms by which Mycobacterium tuberculosis, a slow growing human pathogen, responds to different types of replication stress and DNA damage are unclear. Here, we show that M. tuberculosis RecG rescues E. coli Delta recG cells from replicative stress. The purified M. tuberculosis RecG (MtRecG) and RuvAB(MtRuvAB) proteins catalyze fork reversal of model replication fork structures with and without a leading strand single-stranded DNA gap. Interestingly, single-stranded DNA-binding protein suppresses the MtRecG- and MtRuvAB-mediated fork reversal with substrates that contain lagging strand gap. Notably, our comparative studies with fork structures containing template damage and template switching mechanism of lesion bypass reveal that MtRecG but not MtRuvAB or MtRecA is proficient in driving the fork reversal. Finally, unlike MtRuvAB, we find that MtRecG drives efficient reversal of forks when fork structures are tightly bound by protein. These results provide direct evidence and valuable insights into the underlying mechanism of MtRecG-catalyzed replication fork remodeling and restart pathways in vivo.

Formato

application/pdf

Identificador

http://eprints.iisc.ernet.in/52634/1/Jou_of_Bio_Che_290-40_24119_2015.pdf

Thakur, Roshan Singh and Basavaraju, Shivakumar and Khanduja, Jasbeer Singh and Muniyappa, K and Nagaraju, Ganesh (2015) Mycobacterium tuberculosis RecG Protein but Not RuvAB or RecA Protein Is Efficient at Remodeling the Stalled Replication Forks IMPLICATIONS FOR MULTIPLE MECHANISMS OF REPLICATION RESTART IN MYCOBACTERIA. In: JOURNAL OF BIOLOGICAL CHEMISTRY, 290 (40). pp. 24119-24139.

Publicador

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Relação

http://dx.doi.org/10.1074/jbc.M115.671164

http://eprints.iisc.ernet.in/52634/

Palavras-Chave #Biochemistry
Tipo

Journal Article

PeerReviewed