TY - JOUR
T1 - Knockouts of RecA-Like proteins RadC1 and RadC2 have distinct responses to DNA damage agents in Sulfolobus islandicus
AU - Liang, Peng-Juan
AU - Han, Wenyuan
AU - Huang, Qi-Hong
AU - Li, Yan-Ze
AU - Ni, Jin-Feng
AU - She, Qunxin
AU - Shen, Yu-Long
PY - 2013/10/20
Y1 - 2013/10/20
N2 - RecA family recombinases play essential roles in maintaining genome integrity. A group of RecA-like proteins named RadC are present in all archaea, but their invivo functions remain unclear. In this study, we performed phylogenetic and genetic analysis of two RadC proteins from Sulfolobus islandicus. RadC is closer to the KaiC lineage of cyanobacteria and proteobacteria than to the lineage of the recombinases (RecA, RadA, and Rad51) and the recombinase paralogs (e.g., RadB, Rad55, and Rad51B). Using the recently-established S. islandicus genetic system, we constructed deletion and over-expression strains of radC1 and radC2. Deletion of radC1 rendered the cells more sensitive to DNA damaging agents, methyl methanesulfonate (MMS), hydroxyurea (HU), and ultraviolet (UV) radiation, than the wild type, and a δradC1δradC2 double deletion strain was more sensitive to cisplatin and MMS than the δradC1 single deletion mutant. In addition, ectopic expression of His-tagged RadC1 revealed that RadC1 was co-purified with a putative structure-specific nuclease and ATPase, which is highly conserved in archaea. Our results indicate that both RadC1 and RadC2 are involved in DNA repair. RadC1 may play a general or primary role in DNA repair, while RadC2 plays a role in DNA repair in response to specific DNA damages.
AB - RecA family recombinases play essential roles in maintaining genome integrity. A group of RecA-like proteins named RadC are present in all archaea, but their invivo functions remain unclear. In this study, we performed phylogenetic and genetic analysis of two RadC proteins from Sulfolobus islandicus. RadC is closer to the KaiC lineage of cyanobacteria and proteobacteria than to the lineage of the recombinases (RecA, RadA, and Rad51) and the recombinase paralogs (e.g., RadB, Rad55, and Rad51B). Using the recently-established S. islandicus genetic system, we constructed deletion and over-expression strains of radC1 and radC2. Deletion of radC1 rendered the cells more sensitive to DNA damaging agents, methyl methanesulfonate (MMS), hydroxyurea (HU), and ultraviolet (UV) radiation, than the wild type, and a δradC1δradC2 double deletion strain was more sensitive to cisplatin and MMS than the δradC1 single deletion mutant. In addition, ectopic expression of His-tagged RadC1 revealed that RadC1 was co-purified with a putative structure-specific nuclease and ATPase, which is highly conserved in archaea. Our results indicate that both RadC1 and RadC2 are involved in DNA repair. RadC1 may play a general or primary role in DNA repair, while RadC2 plays a role in DNA repair in response to specific DNA damages.
KW - RecA-like protein
KW - RadA
KW - RadC
KW - Sulfolobus
KW - DNA repair
U2 - 10.1016/j.jgg.2013.05.004
DO - 10.1016/j.jgg.2013.05.004
M3 - Journal article
SN - 1673-8527
VL - 40
SP - 533
EP - 542
JO - Journal of Genetics and Genomics
JF - Journal of Genetics and Genomics
IS - 10
ER -