Mitogen-activated protein kinase phosphatase PHS1 is retained in the cytoplasm by nuclear extrusion signal-dependent and independent mechanisms.
The organization of plant microtubule arrays is thought to be regulated by phosphorylation and other signaling cascades, but the molecular components involved are largely unknown. We have previously found that a dominant missense mutation (phs1-1) in a putative kinase-docking motif of an Arabidopsis PHS1 phosphatase, which belongs to the mitogen-activated protein kinase phosphatase (MKP) family, compromises the stability of cortical microtubules. We here report that suppressor screening of phs1-1 recovered several intragenic recessive mutations in PHS1. In contrast to our previous report, null alleles of PHS1 were almost indistinguishable from the wild type in morphology, but their roots skewed to the abnormal direction when grew in the presence of low doses of a microtubule-destabilizing drug. PHS1 is mainly expressed in elongating cells, where the protein was distributed in the cytoplasm, predominantly in a microsomal fraction. Recruitment of green fluorescent protein-tagged PHS1 in endomembrane aggregates after treatment with brefeldin A or in an endomembrane-organization mutant suggests that an association with endomembranes retains PHS1 in the cytoplasm. A nuclear export signal identified in the C-terminal tail also contributes to the robust cytoplasmic retention of PHS1.
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0192 Nara, Japan.
Jaromir
J
Pytela
Takehide
T
Kato
Takashi
T
Hashimoto
D000483:Alleles; D017360:Arabidopsis / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism*; D002467:Cell Nucleus / Q000201:enzymology*; D005966:Glucuronidase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D049790:Nuclear Export Signals; D010641:Phenotype; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology
D029681:Arabidopsis Proteins; D049790:Nuclear Export Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C514675:PHS1 protein, Arabidopsis; D017027:Protein Tyrosine Phosphatases; D005966:Glucuronidase