Acetylation dependent functions of Rab22a-NeoF1 Fusion Protein in Osteosarcoma.

Theranostics
Xiaoting LiangTiebang Kang

Abstract

Background: Rab22a-NeoF1 fusion gene containing the 1-38aa of Rab22a (Rab22a1-38) plays a decisive role in driving tumor metastasis by activating RhoA via binding to SmgGDS607. However, its intercellular regulation remains unknown. Methods: The Lys7 (K7) acetylation of Rab22a-NeoF1 was initially identified by mass spectrum. Co-transfection, immunoprecipitation and Western blotting were used to characterize the acetyltransferases and deacetylases responsible for the K7 acetylation of Rab22a-NeoF1, and to define the interaction of proteins. The specificity of K7 acetylation of Rab22a-NeoF1 was determined by its specific anti-K7ac-Rab22a-NeoF1 antibody and its K7R mutant. RhoA-GTP was measured by RhoA activation assay. The migration and invasion were assessed by Transwell assay without and with Matrigel matrix, respectively. The orthotopic osteosarcoma metastasis model in vivo was used to monitor the lung metastases of U2OS/MTX300-Luc stably expressing Vector, Rab22a-NeoF1 or its K7R mutant with or without C646, a relatively specific inhibitor of p300/CBP. The unpaired Student t test was used for the statistical significance. Results: The K7 of Rab22a-NeoF1 is acetylated by p300/CBP while is de-acetylated by both HDAC6 and SIRT1. ...Continue Reading

Citations

Aug 26, 2020·Signal Transduction and Targeted Therapy·Kai XieYongguang Tao
Jul 30, 2021·Frontiers in Pharmacology·Xueming XuLi Tian

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Methods Mentioned

BETA
GTPase
ubiquitination
acetylation
Pull-down
Assay
transfection

Software Mentioned

SPSS
GraphPad
GraphPad Prism

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