营养压力（即限制乙酰辅酶A从葡萄糖转化或乙酸酯获取），引起ACSS2在S659位点磷酸化并向核内转移。与TFEB结合的ACSS2会在溶酶体和自噬相关基因启动子区域与染色体结合,促进吸收、利用组蛋白乙酰化循环释放的乙酸脂合成乙酰辅酶A，后者驱动H3组蛋白乙酰化并活化上述基因表达。上述过程促进溶酶体生长，自噬以及脑肿瘤生长。



Figure1.ACSS2S659PhosphorylationbyAMPKInducesNuclearTranslocationofACSS2
 
(B–G)Immunoblotanalyseswereperformedwiththeindicatedantibodies.
 

• U87cellsweredeprivedofglucosefor1hr.Immunofluorescentanalyseswereperformedwithananti-ACSS2antibody.DAPI(blue)wasusedtostainthenuclei.

• U87cellsweredeprivedofglucosefor1hr.Totalcelllysatesandcytosolicandnuclearfractionswereprepared.ActivationofAMPKisreflectedbyAMPKaT172phosphorylation.

• WTandAMPKa1andAMPKa2double-knockout(DKO)MEFsweretreatedwith2-DG(25mM)fortheindicatedperiods.Totalcelllysatesandcytosolicandnuclearfractionswereprepared.

• U87cellsweredeprivedofglucosefor10min.Immunoprecipitationwithananti-AMPKaantibodywasperformed.

• PurifiedAMPKwasmixedwiththeindicatedbacteriallypurifiedHis-ACSS2proteinsand[g-³²P]ATPinthepresenceorabsenceofAMP(100mM).Auto-rADIographicanalysiswasperformed.

• U87cellsexpressingtheindicatedFLAG-ACSS2proteinsweredeprivedofglucosefor1hr.Immunoprecipitationwithananti-FLAGantibodywasperformed.ActivationofAMPKisreflectedbyitssubstrateacetyl-CoAcarboxylase(ACC)S79phosphorylation.

• WTandAMPKa1/2DKOMEFsweredeprivedofglucosefor1hr.Totalcelllysateswereprepared.

 
原文参考链接 

Lietal.,2017,MolecularCell66,1–14June1,2017ª2017ElsevierInc.

本文应用核心抗体