C3G dynamically associates with nuclear speckles and regulates mRNA splicing

Molecular Biology of the Cell
Dhruv Kumar ShakyawarVegesna Radha

Abstract

C3G (Crk SH3 domain binding guanine nucleotide releasing factor) (Rap guanine nucleotide exchange factor 1), essential for mammalian embryonic development, is ubiquitously expressed and undergoes regulated nucleocytoplasmic exchange. Here we show that C3G localizes to SC35-positive nuclear speckles and regulates splicing activity. Reversible association of C3G with speckles was seen on inhibition of transcription and splicing. C3G shows partial colocalization with SC35 and is recruited to a chromatin and RNase-sensitive fraction of speckles. Its presence in speckles is dependent on intact cellular actin cytoskeleton and is lost on expression of the kinase Clk1. Rap1, a substrate of C3G, is also present in nuclear speckles, and inactivation of Rap signaling by expression of GFP-Rap1GAP alters speckle morphology and number. Enhanced association of C3G with speckles is seen on glycogen synthase kinase 3 beta inhibition or differentiation of C2C12 cells to myotubes. CRISPR/Cas9-mediated knockdown of C3G resulted in altered splicing activity of an artificial gene as well as endogenous CD44. C3G knockout clones of C2C12 as well as MDA-MB-231 cells showed reduced protein levels of several splicing factors compared with control cells. ...Continue Reading

References

Nov 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·M R Lerner, J A Steitz
Dec 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·G R ScreatonJ I Bell
Apr 1, 1992·The Journal of Cell Biology·M Carmo-FonsecaA I Lamond
May 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·E A LernerJ A Steitz
Apr 12, 1994·Proceedings of the National Academy of Sciences of the United States of America·S TanakaK Nagashima
Jan 1, 1993·Annual Review of Cell Biology·D L Spector
Feb 1, 1994·The Journal of Cell Biology·R T O'KeefeD L Spector
May 29, 1997·Nature·T MisteliD L Spector
Jul 25, 1997·The Journal of Biological Chemistry·T GotohS Hattori
May 13, 1999·The Journal of Biological Chemistry·T IchibaM Matsuda
Apr 25, 2000·The Journal of Biological Chemistry·N MochizukiM Matsuda
May 12, 2000·Journal of Structural Biology·P J Mintz, D L Spector
Mar 29, 2001·Proceedings of the National Academy of Sciences of the United States of America·J Eilbracht, M S Schmidt-Zachmann
Oct 23, 2001·Methods in Enzymology·Kendall D Carey, Phillip J S Stork
Nov 6, 2001·Molecular Biology of the Cell·M DenegriG Biamonti
Aug 29, 2002·The Journal of Biological Chemistry·Jingwei Meng, Patrick J Casey
Sep 13, 2002·Nature·Zhaolan ZhouRobin Reed
Jun 12, 2003·Experimental Cell Research·Giovanna TabelliniAlberto M Martelli
Aug 19, 2003·Nature Reviews. Molecular Cell Biology·Angus I Lamond, David L Spector
Sep 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sharon Wald KraussRebecca Heald
Feb 6, 2004·The Journal of Cell Biology·Trever G BivonaMark R Philips
Jun 1, 2004·Molecular Biology of the Cell·Noriko SaitohDavid L Spector
Jul 16, 2004·Molecular and Cellular Biology·Catherine HoganYasuyuki Fujita
Dec 6, 2005·Trends in Cell Biology·Korie E Handwerger, Joseph G Gall
May 10, 2006·International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society·K OkinoT Takeshita
Jun 9, 2006·The Anatomical Record. Part A, Discoveries in Molecular, Cellular, and Evolutionary Biology·Lisa L HallJeanne B Lawrence
May 4, 2007·Experimental Cell Research·Vegesna RadhaGhanshyam Swarup
Nov 6, 2007·The Journal of Biological Chemistry·Jos L J van der VeldenRamon C J Langen
Nov 21, 2007·The Journal of Biological Chemistry·Stefan Stamm
Feb 8, 2008·Molecular Biology of the Cell·Véronique BaldinOlivier Coux
Oct 2, 2008·The Journal of Biological Chemistry·Kristine O'BrienMelissa J Moore
Nov 4, 2008·Nature·Eric T WangChristopher B Burge

❮ Previous
Next ❯

Citations

Jan 16, 2021·Biochimica Et Biophysica Acta. Molecular Cell Research·Divya SriramVegesna Radha
Nov 4, 2020·Scientific Reports·Divya SriramVegesna Radha
Feb 25, 2021·Stem Cell Reviews and Reports·Vijay V VishnuP Chandra Shekar

❮ Previous
Next ❯

Methods Mentioned

BETA
nucleotide exchange
GTPases
nuclear translocation
PCR
GTPase
transfections

Software Mentioned

ImageJ
Leica Application Suite
Adobe Photoshop

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.