Non-visual arrestins are constitutively associated with the centrosome and regulate centrosome function.
In addition to regulating receptor activity, non-visual arrestins function as scaffolds for numerous intracellular signaling cascades and as regulators of gene transcription. Here we report that the two non-visual arrestins, arrestin2 and arrestin3, localize to the centrosome, a key organelle involved in microtubule nucleation and bipolar mitotic spindle assembly. Both arrestins co-localized with the centrosomal marker gamma-tubulin during interphase and mitosis and were found in purified centrosome preparations. In vitro binding assays demonstrated that both arrestins directly interact with gamma-tubulin. Knockdown of either arrestin by RNA interference resulted in multinucleation, centrosome amplification, and mitotic defects, although only the loss of arrestin2 triggered aberrant microtubule nucleation. Importantly, overexpression of wild type arrestin rescued the multinucleation phenotype and restored normal centrosome number in arrestin siRNA-transfected cells. Moreover, overexpression of arrestin2 or -3 rescued the multinucleation defect observed in MDA-MB-231 breast cancer cells. Taken together, our data reveal that non-visual arrestins are novel centrosomal components and regulate normal centrosome function.
Shankar
Haripriya
H
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Michal
Allison
A
Kern
Ronald C
RC
Kang
Dong Soo
DS
Gurevich
Vsevolod V
VV
Benovic
Jeffrey L
JL
D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D001943:Breast Neoplasms; D002417:Cattle; D018385:Centrosome / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D007399:Interphase / Q000502:physiology; D007668:Kidney / Q000166:cytology; D018613:Microscopy, Confocal; D008938:Mitosis / Q000502:physiology; D010714:Phosphatidylethanolamines; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D014404:Tubulin / Q000378:metabolism*; D000071557:beta-Arrestins
D019390:Arrestins; D010714:Phosphatidylethanolamines; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; D014404:Tubulin; C082333:arrestin3; D000071557:beta-Arrestins; C100004:transfectACE