Phospholipase C beta 1 is linked to RNA interference of specific genes through translin-associated factor X

Article

关键词: C BETA-1;    EXPRESSION;    BRAIN;    TRAX;    MICE;    ACTIVATION;    COMPLEX;    ROLES;    MODEL;    C3PO;   

DOI  :  10.1096/fj.12-213934

来源: SCIE

【 摘 要 】

Phospholipase C beta 1 (PLC beta 1) is a G-protein-regulated enzyme whose activity results in proliferative and mitogenic changes in the cell. We have previously found that in solution PLC beta 1 binds to the RNA processing protein translin-associated factor X (TRAX) with nanomolar affinity and that this binding competes with G proteins. Here, we show that endogenous PLC beta 1 and TRAX interact in SK-N-SH cells and also in HEK293 cells induced to overexpress PLC beta 1. In HEK293 cells, TRAX overexpression ablates Ca2+ signals generated by G protein-PLC beta 1 activation. TRAX plays a key role in down-regulation of proteins by small, interfering RNA, and PLC beta 1 overexpression completely reverses the 2- to 4-fold down-regulation of GAPDH by siRNA in HEK293 and HeLa cells as seen by an similar to 4-fold recovery in both the transcript and protein levels. Also, down-regulation of endogenous PLC beta 1 in HEK293 and HeLa cells allows for an similar to 20% increase in siRNA(GAPDH) silencing. While PLC beta 1 overexpression results in a 50% reversal of cell death caused by siRNA(LDH), it does not affect cell survival or silencing of other genes (e. g., cyclophilin, Hsp90, translin). PLC beta 1 overexpression in HEK293 and HeLa cells causes a 30% reduction in the total amount of small RNAs. LDH and GAPDH are part of a complex that promotes H2B synthesis that allows cells to progress through the S phase. We find that PLC beta 1 reverses the cell death and completely rescues H2B levels caused by siRNA knockdown of LDH or GAPDH. Taken together, our study shows a novel role of PLC beta 1 in gene regulation through TRAX association.-Philip, F., Guo, Y., Aisiku, O., Scarlata, S. Phospholipase C beta 1 is linked to RNA interference of specific genes through translin-associated factor X. FASEB J. 26, 4903-4913 (2012). www.fasebj.org

【 授权许可】

Free