A mutation in MRH2 kinesin enhances the root hair tip growth defect caused by constitutively activated ROP2 small GTPase in Arabidopsis.
Root hair tip growth provides a unique model system for the study of plant cell polarity. Transgenic plants expressing constitutively active (CA) forms of ROP (Rho-of-plants) GTPases have been shown to cause the disruption of root hair polarity likely as a result of the alteration of actin filaments (AF) and microtubules (MT) organization. Towards understanding the mechanism by which ROP controls the cytoskeletal organization during root hair tip growth, we have screened for CA-rop2 suppressors or enhancers using CA1-1, a transgenic line that expresses CA-rop2 and shows only mild disruption of tip growth. Here, we report the characterization of a CA-rop2 enhancer (cae1-1 CA1-1) that exhibits bulbous root hairs. The cae1-1 mutation on its own caused a waving and branching root hair phenotype. CAE1 encodes the root hair growth-related, ARM domain-containing kinesin-like protein MRH2 (and thus cae1-1 was renamed to mrh2-3). Cortical MT displayed fragmentation and random orientation in mrh2 root hairs. Consistently, the MT-stabilizing drug taxol could partially rescue the wavy root hair phenotype of mrh2-3, and the MT-depolymerizing drug Oryzalin slightly enhanced the root hair tip growth defect in CA1-1. Interestingly, the addition of the actin-depolymerizing drug Latrunculin B further enhanced the Oryzalin effect. This indicates that the cross-talk of MT and AF organization is important for the mrh2-3 CA1-1 phenotype. Although we did not observe an apparent effect of the MRH2 mutation in AF organization, we found that mrh2-3 root hair growth was more sensitive to Latrunculin B. Moreover, an ARM domain-containing MRH2 fragment could bind to the polymerized actin in vitro. Therefore, our genetic analyses, together with cell biological and pharmacological evidence, suggest that the plant-specific kinesin-related protein MRH2 is an important component that controls MT organization and is likely involved in the ROP2 GTPase-controlled coordination of AF and MT during polarized growth of root hairs.
Yang
Guohua
G
Department of Biological Sciences, Lehman College, City University of New York, Bronx, New York, United States of America.
Gao
Peng
P
Zhang
Hua
H
Huang
Shanjin
S
Zheng
Zhi-Liang
ZL
D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000502:physiology; D017931:DNA Primers / Q000737:chemistry; D019204:GTP-Binding Proteins / Q000235:genetics*; D005816:Genetic Complementation Test; D049452:Green Fluorescent Proteins / Q000737:chemistry; D016547:Kinesins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D018519:Meristem / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D008957:Models, Genetic; D020559:Monomeric GTP-Binding Proteins / Q000737:chemistry*; D009154:Mutation; D018517:Plant Roots / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D029681:Arabidopsis Proteins; D017931:DNA Primers; D019204:GTP-Binding Proteins; D049452:Green Fluorescent Proteins; D016547:Kinesins; D020559:Monomeric GTP-Binding Proteins; D011993:Recombinant Fusion Proteins; C529971:MRH2 protein, Arabidopsis; C499714:AT1G20090 protein, Arabidopsis