Functional analysis of the Volvox carteri asymmetric division protein GlsA.
The Zuotin-family J protein chaperone GlsA is essential for the asymmetric divisions that establish germ and somatic cell initials during embryogenesis in the green alga Volvox carteri, but it is not known on what cellular process GlsA acts to carry out this function. Most GlsA protein is nuclear, and GlsA possesses two SANT domains, suggesting that GlsA may function as a transcriptional regulator. On the other hand, close homologs from yeast and mice are ribosome-associated factors that regulate translation fidelity, implying GlsA might also regulate translation. Here we set out to gain additional evidence regarding the function of GlsA, specifically with respect to its possible involvement in transcription and translation. We found that like zuotin mutants, glsA mutants are ultrasensitive to both cold and to the ribosome-binding aminoglycoside antibiotic paromomycin, so some fraction of GlsA is likely to be ribosome associated. We also found that GlsA co-immunoprecipitates with histones and that this interaction is dependent on the presence of intact SANT domains. Through rescue experiments using transgenes that encode GlsA variants, we determined that the growth and asymmetric division defects of the glsA mutant are separable-a GlsA variant that rescued the growth defects did not completely rescue the asymmetric division phenotype. Considered in total, our results suggest that GlsA acts both at the level of translation and transcription, but the function that is essential for tolerance to paromomycin and cold is not sufficient for asymmetric cell division.
Pappas
Valeria
V
Department of Biological Sciences, University of Maryland, Baltimore County, Baltimore, MD 21250, USA.
Miller
Stephen M
SM
D020418:Algal Proteins / Q000502:physiology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D005075:Biological Evolution; D002455:Cell Division / Q000502:physiology*; D017931:DNA Primers; D006360:Heat-Shock Proteins / Q000502:physiology*; D047468:Immunoprecipitation; D010303:Paromomycin / Q000494:pharmacology; D044446:Volvox / Q000235:genetics / Q000502:physiology*
D020418:Algal Proteins; D000900:Anti-Bacterial Agents; D017931:DNA Primers; D006360:Heat-Shock Proteins; D010303:Paromomycin