The Journal of Cell Biology, Vol 135, 1853-1865, Copyright © 1996 by The Rockefeller University Press

ARTICLES

The sup-pf-2 mutations of Chlamydomonas alter the activity of the outer dynein arms by modification of the gamma-dynein heavy chain

G Rupp, E O'Toole, LC Gardner, BF Mitchell and ME Porter
Department of Cell Biology and Neuroanatomy, University of Minnesota Medical School, Minneapolis 55455, USA.

The sup-pf-2 mutation is a member of a group of dynein regulatory mutations that are capable of restoring motility to paralyzed central pair or radial spoke defective strains. Previous work has shown that the flagellar beat frequency is reduced in sup-pf-2, but little else was known about the sup-pf-2 phenotype (Huang, B., Z. Ramanis, and D.J.L. Luck. 1982. Cell. 28:115-125; Brokaw, C.J., and D.J.L. Luck. 1985. Cell Motil. 5:195-208). We have reexamined sup-pf-2 using improved biochemical and structural techniques and by the analysis of additional sup-pf-2 alleles. We have found that the sup-pf-2 mutations are associated with defects in the outer dynein arms. Biochemical analysis of sup-pf-2-1 axonemes indicates that both axonemal ATPase activity and outer arm polypeptides are reduced by 40-50% when compared with wild type. By thin-section EM, these defects correlate with an approximately 45% loss of outer dynein arm structures. Interestingly, this loss is biased toward a subset of outer doublets, resulting in a radial asymmetry that may reflect some aspect of outer arm assembly. The defects in outer arm assembly do not appear to result from defects in either the outer doublet microtubules or the outer arm docking structures, but rather appear to result from defects in outer dynein arm components. Analysis of new sup-pf-2 mutations indicates that the severity of the outer arm assembly defects varies with different alleles. Complementation tests and linkage analysis reveal that the sup- pf-2 mutations are alleles of the PF28/ODA2 locus, which is thought to encode the gamma-dynein heavy chain subunit of the outer arm. The sup- pf-2 mutations therefore appear to alter the activity of the outer dynein arms by modification of the gamma-dynein heavy chain.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Liu, Z., Takazaki, H., Nakazawa, Y., Sakato, M., Yagi, T., Yasunaga, T., King, S. M., Kamiya, R. (2008). Partially Functional Outer-Arm Dynein in a Novel Chlamydomonas Mutant Expressing a Truncated {gamma} Heavy Chain. Eukaryot Cell 7: 1136-1145 [Abstract] [Full Text]  
• Sakato, M., Sakakibara, H., King, S. M. (2007). Chlamydomonas Outer Arm Dynein Alters Conformation in Response to Ca2+. Mol. Biol. Cell 18: 3620-3634 [Abstract] [Full Text]  
• Ralston, K. S., Lerner, A. G., Diener, D. R., Hill, K. L. (2006). Flagellar Motility Contributes to Cytokinesis in Trypanosoma brucei and Is Modulated by an Evolutionarily Conserved Dynein Regulatory System.. Eukaryot Cell 5: 696-711 [Abstract] [Full Text]  
• Wiley, L. A., Rupp, G. R., Steinle, J. J. (2005). Sympathetic Innervation Regulates Basement Membrane Thickening and Pericyte Number in Rat Retina. IOVS 46: 744-748 [Abstract] [Full Text]  
• Rupp, G., Porter, M. E. (2003). A subunit of the dynein regulatory complex in Chlamydomonas is a homologue of a growth arrest-specific gene product. JCB 162: 47-57 [Abstract] [Full Text]  
• Kispert, A, Petry, M, Olbrich, H, Volz, A, Ketelsen, U-P, Horvath, J, Melkaoui, R, Omran, H, Zariwala, M, Noone, P G, Knowles, M (2003). Genotype-phenotype correlations in PCD patients carrying DNAH5 mutations. Thorax 58: 552-554 [Full Text]  
• Wargo, M. J., Smith, E. F. (2003). Asymmetry of the central apparatus defines the location of active microtubule sliding in Chlamydomonas flagella. Proc. Natl. Acad. Sci. USA 100: 137-142 [Abstract] [Full Text]  
• Pennarun, G., Bridoux, A.-M., Escudier, E., Dastot-Le Moal, F., Cacheux, V., Amselem, S., Duriez, B. (2002). Isolation and Expression of the Human hPF20 Gene Orthologous to Chlamydomonas pf20 . Evaluation as a Candidate for Axonemal Defects of Respiratory Cilia and Sperm Flagella. Am. J. Respir. Cell Mol. Bio. 26: 362-370 [Abstract] [Full Text]  
• Rupp, G., O'Toole, E., Porter, M. E. (2001). The Chlamydomonas PF6 Locus Encodes a Large Alanine/Proline-Rich Polypeptide That Is Required for Assembly of a Central Pair Projection and Regulates Flagellar Motility. Mol. Biol. Cell 12: 739-751 [Abstract] [Full Text]  
• Porter, M. E., Sale, W. S. (2000). The 9 + 2 Axoneme Anchors Multiple Inner Arm Dyneins and a Network of Kinases and Phosphatases That Control Motility. JCB 151: 37-42 [Full Text]  
• Omran, H., Häffner, K., Völkel, A., Kuehr, J., Ketelsen, U.-P., Ross, U.-H., Konietzko, N., Wienker, T., Brandis, M., Hildebrandt, F. (2000). Homozygosity Mapping of a Gene Locus for Primary Ciliary Dyskinesia on Chromosome 5p and Identification of the Heavy Dynein Chain DNAH5 as a Candidate Gene. Am. J. Respir. Cell Mol. Bio. 23: 696-702 [Abstract] [Full Text]  
• Yang, P, Fox, L, Colbran, R., Sale, W. (2000). Protein phosphatases PP1 and PP2A are located in distinct positions in the Chlamydomonas flagellar axoneme. J. Cell Sci. 113: 91-102 [Abstract]  
• Myster, S. H., Knott, J. A., Wysocki, K. M., O'Toole, E., Porter, M. E. (1999). Domains in the 1{alpha} Dynein Heavy Chain Required for Inner Arm Assembly and Flagellar Motility in Chlamydomonas. JCB 146: 801-818 [Abstract] [Full Text]  
• Mitchell, D. R., Sale, W. S. (1999). Characterization of a Chlamydomonas Insertional Mutant that Disrupts Flagellar Central Pair Microtubule-associated Structures. JCB 144: 293-304 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents