TY - JOUR

T1 - Src-mediated phosphorylation of GAPDH regulates its nuclear localization and cellular response to DNA damage

AU - Ci, Shusheng

AU - Xia, Wen

AU - Liang, Weichu

AU - Qin, Lihong

AU - Zhang, Yilan

AU - Dianov, Grigory L.

AU - Wang, Meina

AU - Zhao, Xingqi

AU - Wu, Congye

AU - Alagamuthu, Karthick Kumar

AU - Hu, Zhigang

AU - He, Lingfeng

AU - Pan, Feiyan

AU - Guo, Zhigang

N1 - Publisher Copyright: © 2020 Federation of American Societies for Experimental Biology Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2020/8/1

Y1 - 2020/8/1

N2 - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme involved in energy metabolism. Recently, GAPDH has been suggested to have extraglycolytic functions in DNA repair, but the underlying mechanism for the GAPDH response to DNA damage remains unclear. Here, we demonstrate that the tyrosine kinase Src is activated under DNA damage stress and phosphorylates GAPDH at Tyr41. This phosphorylation of GAPDH is essential for its nuclear translocation and DNA repair function. Blocking the nuclear import of GAPDH by suppressing Src signaling or through a GAPDH Tyr41 mutation impairs its response to DNA damage. Nuclear GAPDH is recruited to DNA lesions and associates with DNA polymerase β (Pol β) to function in DNA repair. Nuclear GAPDH promotes Pol β polymerase activity and increases base excision repair (BER) efficiency. Furthermore, GAPDH knockdown dramatically decreases BER efficiency and sensitizes cells to DNA damaging agents. Importantly, the knockdown of GAPDH in colon cancer SW480 cells and xenograft models effectively enhances their sensitivity to the chemotherapeutic drug 5-FU. In summary, our findings provide mechanistic insight into the new function of GAPDH in DNA repair and suggest a potential therapeutic target in chemotherapy.

AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme involved in energy metabolism. Recently, GAPDH has been suggested to have extraglycolytic functions in DNA repair, but the underlying mechanism for the GAPDH response to DNA damage remains unclear. Here, we demonstrate that the tyrosine kinase Src is activated under DNA damage stress and phosphorylates GAPDH at Tyr41. This phosphorylation of GAPDH is essential for its nuclear translocation and DNA repair function. Blocking the nuclear import of GAPDH by suppressing Src signaling or through a GAPDH Tyr41 mutation impairs its response to DNA damage. Nuclear GAPDH is recruited to DNA lesions and associates with DNA polymerase β (Pol β) to function in DNA repair. Nuclear GAPDH promotes Pol β polymerase activity and increases base excision repair (BER) efficiency. Furthermore, GAPDH knockdown dramatically decreases BER efficiency and sensitizes cells to DNA damaging agents. Importantly, the knockdown of GAPDH in colon cancer SW480 cells and xenograft models effectively enhances their sensitivity to the chemotherapeutic drug 5-FU. In summary, our findings provide mechanistic insight into the new function of GAPDH in DNA repair and suggest a potential therapeutic target in chemotherapy.

KW - base excision repair

KW - chemotherapy

KW - DNA damage

KW - GAPDH

KW - Pol β

KW - Src

KW - ACTIVATION

KW - PROTEIN

KW - BETA-GENE MUTATION

KW - KINASE

KW - POLYMERASE-BETA

KW - Pol beta

KW - CANCER

KW - BASE EXCISION-REPAIR

KW - RESISTANCE

KW - EXPRESSION

KW - HUMAN GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE

UR - http://www.scopus.com/inward/record.url?scp=85086446829&partnerID=8YFLogxK

U2 - 10.1096/fj.201902904RR

DO - 10.1096/fj.201902904RR

M3 - Article

C2 - 32539222

AN - SCOPUS:85086446829

VL - 34

SP - 10443

EP - 10461

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 8

ER -