An in vitro study on mechanism of ANP32B in the promotion of tumors

doi:10.16150/j.1671-2870.2018.02.013

Abstract

Abstract: Objective: To investigate the molecular mechanism of ANP32B （acidic leucine-rich nuclear phosphoprotein 32B）in the promotion of tumors. Methods: ANP32B interaction proteins were searched and identified by immunoprecipitation(IP)-coupled LC-MS/MS technology. Immunofluorescence was used to identify the localization of ANP32B and karyopherin α6 (KPNA6). KPNA6 was knockdown by shRNA. GST-pulldown assay was used to identify the direct interaction of ANP32B and KPNA6 and domains of interaction. Results: Immunoprecipitation(IP)-coupled LC-MS/MS technology identified the interaction of KPNA6 with ANP32B, and the N-terminal 1-161 amino acid of ANP32B was not required for its interaction with KPNA6. In vitro GST-pulldown assay demonstrated the direct interaction between KPNA6 and ANP32B, and the nuclear localization signal (NLS) of ANP32B was essential for its interaction with KPNA6. Inhibition of KPNA6 expression by shRNA had no effect on the nuclear localization of ANP32B. Conclusions: ANP32B interactes with KPNA6 via its NLS domain, and ANP32B is co-localized with KPNA6 in the nucleus. Knockdown of KPNA6 does not influence the nuclear localization of ANP32B.

Key words: Acidic leucine-rich nuclear phosphoprotein 32B, Karyopherin α6, Interaction, BT594 cell line

CLC Number:

R730.1

ZHU Xiaona, YANG Shuo, HE Ping, ZHANG Huilin, FENG Ru, YU Yun. An in vitro study on mechanism of ANP32B in the promotion of tumors[J]. Journal of Diagnostics Concepts & Practice, 2018, 17(02): 186-190.

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