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2015 ; 290
(24
): 14826-40
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Genome Protection by the 9-1-1 Complex Subunit HUS1 Requires Clamp Formation, DNA
Contacts, and ATR Signaling-independent Effector Functions
#MMPMID25911100
Lim PX
; Patel DR
; Poisson KE
; Basuita M
; Tsai C
; Lyndaker AM
; Hwang BJ
; Lu AL
; Weiss RS
J Biol Chem
2015[Jun]; 290
(24
): 14826-40
PMID25911100
show ga
The RAD9A-HUS1-RAD1 (9-1-1) complex is a heterotrimeric clamp that promotes
checkpoint signaling and repair at DNA damage sites. In this study, we elucidated
HUS1 functional residues that drive clamp assembly, DNA interactions, and
downstream effector functions. First, we mapped a HUS1-RAD9A interface residue
that was critical for 9-1-1 assembly and DNA loading. Next, we identified
multiple positively charged residues in the inner ring of HUS1 that were crucial
for genotoxin-induced 9-1-1 chromatin localization and ATR signaling. Finally, we
found two hydrophobic pockets on the HUS1 outer surface that were important for
cell survival after DNA damage. Interestingly, these pockets were not required
for 9-1-1 chromatin localization or ATR-mediated CHK1 activation but were
necessary for interactions between HUS1 and its binding partner MYH, suggesting
that they serve as interaction domains for the recruitment and coordination of
downstream effectors at damage sites. Together, these results indicate that, once
properly loaded onto damaged DNA, the 9-1-1 complex executes multiple, separable
functions that promote genome maintenance.