Cortical localization of the Gα protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division

Katayoun Afshar, Francis S. Willard, Kelly Colombo, David P. Siderovski, Pierre Gönczy

Research output: Contribution to journalArticle

63 Scopus citations

Abstract

Understanding of the mechanisms governing spindle positioning during asymmetric division remains incomplete. During unequal division of one-cell stage C. elegans embryos, the Gα proteins GOA-1 and GPA-16 act in a partially redundant manner to generate pulling forces along astral microtubules. Previous work focused primarily on GOA-1, whereas the mechanisms by which GPA-16 participates in this process are not well understood. Here, we report that GPA-16 is present predominantly at the cortex of one-cell stage embryos. Using co-immunoprecipitation and surface plasmon resonance binding assays, we find that GPA-16 associates with RIC-8 and GPR-1/2, two proteins known to be required for pulling force generation. Using spindle severing as an assay for pulling forces, we demonstrate that inactivation of the Gβ protein GPB-1 renders GPA-16 and GOA-1 entirely redundant. This suggests that the two Gα proteins can activate the same pathway and that their dual presence is normally needed to counter Gβγ. Using nucleotide exchange assays, we establish that whereas GPR-1/2 acts as a guanine nucleotide dissociation inhibitor (GDI) for GPA-16, as it does for GOA-1, RIC-8 does not exhibit guanine nucleotide exchange factor (GEF) activity towards GPA-16, in contrast to its effect on GOA-1. We establish in addition that RIC-8 is required for cortical localization of GPA-16, whereas it is not required for that of GOA-1. Our analysis demonstrates that this requirement toward GPA-16 is distinct from the known function of RIC-8 in enabling interaction between Gα proteins and GPR-1/2, thus providing novel insight into the mechanisms of asymmetric spindle positioning.

Original languageEnglish
Pages (from-to)4449-4459
Number of pages11
JournalDevelopment
Volume132
Issue number20
DOIs
StatePublished - 1 Oct 2005

Keywords

  • C. elegans embryos
  • G protein
  • Spindle positioning

Fingerprint Dive into the research topics of 'Cortical localization of the Gα protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division'. Together they form a unique fingerprint.

  • Cite this