Laura K Hilton1, Kavisha Gunawardane1, Joo Wan Kim1, Marianne C Schwarz1, Lynne M Quarmby2. 1. Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC V5A 1S6, Canada. 2. Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC V5A 1S6, Canada. Electronic address: quarmby@sfu.ca.
Abstract
BACKGROUND: Many of the diverse functions of cilia depend upon tight control of their length. Steady-state length reflects a balance between rates of ciliary assembly and disassembly, two parameters likely controlled by a length sensor of unknown identity or mechanism. RESULTS: A null mutation in Chlamydomonas CNK2, a member of the evolutionarily conserved family of NIMA-related kinases, reveals feedback regulation of assembly and disassembly rates. cnk2-1 mutant cells have a mild long-flagella (lf) phenotype as a consequence of reduced rates of flagellar disassembly. This is in contrast to the strong lf mutant lf4-7, which exhibits an aberrantly high rate of assembly. Cells carrying both mutations have even longer flagella than lf4-7 single mutants do. In addition to their high rate of assembly, lf4-7 mutants have a CNK2-dependent increase in disassembly rate. Finally, cnk2-1 cells have a decreased rate of turnover of flagellar subunits at the tip of the flagellum, demonstrating that the effects on disassembly are compensated by a reduced rate of assembly. CONCLUSIONS: We propose a model wherein CNK2 and LF4 modulate rates of disassembly and assembly respectively in a feedback loop that is activated when flagella exceed optimal length.
BACKGROUND: Many of the diverse functions of cilia depend upon tight control of their length. Steady-state length reflects a balance between rates of ciliary assembly and disassembly, two parameters likely controlled by a length sensor of unknown identity or mechanism. RESULTS: A null mutation in Chlamydomonas CNK2, a member of the evolutionarily conserved family of NIMA-related kinases, reveals feedback regulation of assembly and disassembly rates. cnk2-1 mutant cells have a mild long-flagella (lf) phenotype as a consequence of reduced rates of flagellar disassembly. This is in contrast to the strong lf mutant lf4-7, which exhibits an aberrantly high rate of assembly. Cells carrying both mutations have even longer flagella than lf4-7 single mutants do. In addition to their high rate of assembly, lf4-7 mutants have a CNK2-dependent increase in disassembly rate. Finally, cnk2-1 cells have a decreased rate of turnover of flagellar subunits at the tip of the flagellum, demonstrating that the effects on disassembly are compensated by a reduced rate of assembly. CONCLUSIONS: We propose a model wherein CNK2 and LF4 modulate rates of disassembly and assembly respectively in a feedback loop that is activated when flagella exceed optimal length.
Authors: William B Ludington; Hiroaki Ishikawa; Yevgeniy V Serebrenik; Alex Ritter; Rogelio A Hernandez-Lopez; Julia Gunzenhauser; Elisa Kannegaard; Wallace F Marshall Journal: Biophys J Date: 2015-03-24 Impact factor: 4.033