Restraint of the G2/M Transition by the SR/RRM family mRNA Shuttling Binding Protein SNXAHRB1 in Aspergillus nidulans
Steven W. James, Travis Banta, James Barra, Lorela Ciraku, Clifford Coile, Zach Cuda, Ryan Day, Cheshil Dixit, Steven Eastlack, Anh Giang, James Goode, Alexis Guice, Yulon Huff, Sara Humbert, Christina Kelliher, Julie Kobie, Emily Kohlbrenner, Faustin Mwambutsa, Amanda Orzechowski, Kristin Shingler, Casey Spell, Sarah Lea Anglin

Abstract

Control of the eukaryotic G2/M transition by CDC2/CYCLINB is tightly regulated by protein-protein interactions, protein phosphorylations, and nuclear localization of CDC2/CYCLINB. We previously reported a screen, in Aspergillus nidulans, for extragenic suppressors of nimX2cdc2 that resulted in the identification of the cold-sensitive snxA1 mutation. We demonstrate here that snxA1 suppresses defects in regulators of the CDK1 mitotic induction pathway, including nimX2cdc2, nimE6cyclinB, and nimT23cdc25, but does not suppress G2-arresting nimA1/nimA5 mutations, the S-arresting nimE10cyclinB mutation or three other G1/S phase mutations. snxA encodes the A. nidulans homolog of Saccharomyces cerevisiae Hrb1/Gbp2, nonessential shuttling mRNA binding proteins belonging to the SR (Serine-Arginine Rich) and RRM (RNA Recognition Motif) protein family, and human hnRNP-M, a spliceosomal component involved in pre-mRNA processing and alternative splicing. snxAHrb1 is nonessential, its deletion phenocopies the snxA1 mutation, and its overexpression rescues snxA1 and ΔsnxA mutant phenotypes. snxA1 and a second allele isolated in this study, snxA2, are hypomorphic mutations that result from decreased transcript and protein levels, suggesting that snxA acts normally to restrain cell cycle progression. SNXAHRB1 is predominantly nuclear, but is not retained in the nucleus during the partially-closed mitosis of A. nidulans. We show that the snxA1 mutation does not suppress nimX2 by altering NIMX2CDC2/NIMECYCLINB kinase activity, and that snxA1 or ΔsnxA alter localization patterns of NIMECYCLINB at the restrictive temperatures for snxA1 and nimX2. Together, these findings suggest a novel and previously unreported role of an SR/RRM family protein in cell cycle regulation, specifically in control of the CDK1 mitotic induction pathway.

  • Received June 18, 2014.
  • Accepted July 31, 2014.