[SapM-induced fusion blocking of autophagosome-lysosome is depended on interaction with Rab7].
Objective To study the role of Rab7 in the blockage of autophagosome-lysosome fusion induced by secretory acid phosphatase (SapM), a virulence factor of mycobacterium tuberculosis. Methods The Raw264.7 cells were transfected with siRab7, and the P62 was detected using Western blotting. After transfected with mCherry-SapM, the co-localization of SapM and Rab7 in Raw264.7 cells was detected by immunofluorescence cytochemistry and the interaction of SapM with Rab7 was determined by co-immunoprecipitation. SapM mutants including SapM(δ ARCA), SapM(δ FRED) and SapM(δ CT) were used to transfect Raw264.7 cells, and their associations with Rab7 were analyzed. Results The treatment of siRab7 induced a significant increase of P62 in these cells. Immunofluorescence cytochemistry showed the intracellular co-localization of SapM and Rab7. Co-immunoprecipitation showed that SapM and Rab7 were precipitated by each other. Only SapM(δ CT) failed to interact with Rab7 among the three SapM mutants. Conclusion The inhibition of autophagosome-lysosome fusion induced by SapM is dependent on the interaction between SapM and Rab7.
Medical Inspection Center, Department of Medical Immunology, Medical School, Huainan 232001, China.
Dong
D
Hu
Medical Inspection Center, Medical School, Huainan 232001, China.
Wan
W
Wang
Medical Inspection Center, Medical School, Huainan 232001, China.
Runpeng
R
Zhao
Medical Inspection Center, Medical School, Huainan 232001, China.
Xuewei
X
Xu
Affiliated Cancer Hospital, Anhui University of Science and Technology, Huainan 232001, China.
Yingru
Y
Xing
Affiliated Cancer Hospital, Anhui University of Science and Technology, Huainan 232001, China.
Shengfa
S
Ni
Affiliated Cancer Hospital, Anhui University of Science and Technology, Huainan 232001, China.
Congjing
C
Xu
Affiliated Cancer Hospital, Anhui University of Science and Technology, Huainan 232001, China.
Baoxian
B
Tie
Medical Inspection Center, Department of Medical Immunology, Medical School, Huainan 232001, China. *Corresponding authors, E-mail:lory456@126.com.
Rongbo
R
Zhang
Medical Inspection Center, Department of Medical Immunology, Medical School, Huainan 232001, China. *Corresponding authors, E-mail: wujing8008@126.com.
Jing
J
Wu
D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000071182:Autophagosomes / Q000201:enzymology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008247:Lysosomes / Q000201:enzymology*; D008264:Macrophages / Q000201:enzymology / Q000382:microbiology; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics; D011485:Protein Binding; D000067996:RAW 264.7 Cells; D014376:Tuberculosis / Q000201:enzymology* / Q000235:genetics / Q000382:microbiology; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000090802:rab7 GTP-Binding Proteins
D001426:Bacterial Proteins; D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; C000716465:rab7 GTP-binding proteins, mouse; D000135:Acid Phosphatase; D020691:rab GTP-Binding Proteins