title: Regulation of the oncoprotein KLF8 by a switch between acetylation and sumoylation.
abstract: KLF8 regulates target genes by recruiting the p300 and PCAF co-activators via glutamines (Q) 118 and 248, the CtBP co-repressor to 86PVDLS90 or SUMO to lysine (K) 67. Here we examined how these interactions coordinate to regulate KLF8 transactivity. Mass spectrometry and immunoprecipitations determined that p300 and/or PCAF promoted KLF8 acetylation at K67, K93, and K95 and this acetylation was abolished in lysine-to-arginine (R) mutants. Treatment with HDAC inhibitors or expression of co-activators inhibited sumoylation at K67. K93R or K95R mutation exerted high levels of sumoylation while the acetylation mimetic mutations K93Q and K95Q blocked the sumoylation. Interestingly, CtBP promoted sumoylation at K67 of wild-type but not AVALF mutant KLF8, and KLF8 interaction with CtBP was inhibited by treatment with the HDAC inhibitors, ectopic expression of the co-activators, or K93Q or K95Q mutation. Promoter reporter assays showed that CtBP inhibited KLF8 transactivity which was rescued by PCAF or p300 expresson. Finally, KLF8-mediated cyclin D1 protein expression and cell cycle progression were significantly decreased in the K93R and K95R but increased in the K93Q, K95Q, K67R or K67Q mutant. Taken together, these results identified a novel mechanism by which co-activators promote KLF8 transactivity by competing with SUMO for K67 modification and by acetylating K93 and K95 to inhibit CtBP-induced K67 sumoylation.
authors.forename: Alison M
authors.initials: AM
authors.lastname: Urvalek
authors.forename: Heng
authors.initials: H
authors.lastname: Lu
authors.forename: Xianhui
authors.initials: X
authors.lastname: Wang
authors.forename: Tianshu
authors.initials: T
authors.lastname: Li
authors.forename: Lin
authors.initials: L
authors.lastname: Yu
authors.forename: Jinghua
authors.initials: J
authors.lastname: Zhu
authors.forename: Qishan
authors.initials: Q
authors.lastname: Lin
authors.forename: Jihe
authors.initials: J
authors.lastname: Zhao