BLM helicase stimulates the ATPase and chromatin-remodeling activities of RAD54.
Mutation of BLM helicase results in the autosomal recessive disorder Bloom syndrome (BS). Patients with BS exhibit hyper-recombination and are prone to almost all forms of cancer. BLM can exhibit its anti-recombinogenic function either by dissolution of double Holliday junctions or by disruption of RAD51 nucleofilaments. We have now found that BLM can interact with the pro-recombinogenic protein RAD54 through an internal ten-residue polypeptide stretch in the N-terminal region of the helicase. The N-terminal region of BLM prevented the formation of RAD51-RAD54 complex, both in vitro and in vivo. Using the fluorescence recovery after photobleaching (FRAP) technique, we found that RAD54 and BLM rapidly and concurrently, yet transiently, bound to the chromatinized foci. Presence of BLM enhanced the mobility of both soluble and chromatinized RAD51 but not RAD54. The BLM-RAD54 interaction could occur even in absence of functional RAD51. The N-terminal 1-212 amino acids of BLM or an ATPase-dead mutant of the full-length helicase enhanced the ATPase and chromatin-remodeling activities of RAD54. These results indicate that apart from its dominant function as an anti-recombinogenic protein, BLM also has a transient pro-recombinogenic function by enhancing the activity of RAD54.
Srivastava
Vivek
V
National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India.
Modi
Priyanka
P
Tripathi
Vivek
V
Mudgal
Richa
R
De
Siddharth
S
Sengupta
Sagar
S
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D002460:Cell Line; D002843:Chromatin / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D042002:Chromatin Assembly and Disassembly; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D006801:Humans; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D053484:RecQ Helicases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D002843:Chromatin; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D051135:Rad51 Recombinase; D000251:Adenosine Triphosphatases; C097151:Bloom syndrome protein; D004265:DNA Helicases; C103532:RAD54L protein, human; D053484:RecQ Helicases