The Chlamydomonas IDA7 locus encodes a 140-kDa dynein intermediate chain required to assemble the I1 inner arm complex

被引:68
作者
Perrone, CA
Yang, PF
O'Toole, E
Sale, WS
Porter, ME [1 ]
机构
[1] Univ Minnesota, Sch Med, Dept Cell Biol & Neuroanat, Minneapolis, MN 55455 USA
[2] Emory Univ, Sch Med, Dept Anat & Cell Biol, Atlanta, GA 30322 USA
[3] Univ Colorado, Dept Mol Cellular & Dev Biol, Boulder, CO 80309 USA
关键词
D O I
10.1091/mbc.9.12.3351
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
To identify new loci that are involved in the assembly and targeting of dynein complexes, we have screened a collection of motility mutants that were generated by insertional mutagenesis. One such mutant, 5B10, lacks the inner arm isoform known as the I1 complex. This isoform is located proximal to the first radial spoke in each 96-nm axoneme repeat and is an important target for the regulation of flagellar motility. Complementation tests reveal that 5B10 represents a new I1 locus, IDA7. Biochemical analyses confirm that ida7 axonemes lack at least five I1 complex subunits. Southern blots probed with a clone containing the gene encoding the 140-kDa intermediate chain (IC) indicate that the ida7 mutation is the result of plasmid insertion into the IC140 gene. Transformation with a wild-type copy of the IC140 gene completely rescues the mutant defects. Surprisingly, transformation with a construct of the IC140 gene lacking the first four exons of the coding sequence also rescues the mutant phenotype. These studies indicate that IC140 is essential for assembly of the I1 complex, but unlike other dynein ICs, the N-terminal region is not critical for its activity.
引用
收藏
页码:3351 / 3365
页数:15
相关论文
共 70 条
[1]   BENDING PATTERNS OF CHLAMYDOMONAS FLAGELLA .4. MUTANTS WITH DEFECTS IN INNER AND OUTER DYNEIN ARMS INDICATE DIFFERENCES IN DYNEIN ARM FUNCTION [J].
BROKAW, CJ ;
KAMIYA, R .
CELL MOTILITY AND THE CYTOSKELETON, 1987, 8 (01) :68-75
[2]   REPEATED CONSENSUS SEQUENCE AND PSEUDOPROMOTERS IN THE 4 COORDINATELY REGULATED TUBULIN GENES OF CHLAMYDOMONAS-REINHARDI [J].
BRUNKE, KJ ;
ANTHONY, JG ;
STERNBERG, EJ ;
WEEKS, DP .
MOLECULAR AND CELLULAR BIOLOGY, 1984, 4 (06) :1115-1124
[3]   SEQUENCES CONTROLLING TRANSCRIPTION OF THE CHLAMYDOMONAS-REINHARDTII BETA(2)-TUBULIN GENE AFTER DEFLAGELLATION AND DURING THE CELL-CYCLE [J].
DAVIES, JP ;
GROSSMAN, AR .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (08) :5165-5174
[4]   THE ARGININOSUCCINATE LYASE GENE OF CHLAMYDOMONAS-REINHARDTII - AN IMPORTANT TOOL FOR NUCLEAR TRANSFORMATION AND FOR CORRELATING THE GENETIC AND MOLECULAR MAPS OF THE ARG7 LOCUS [J].
DEBUCHY, R ;
PURTON, S ;
ROCHAIX, JD .
EMBO JOURNAL, 1989, 8 (10) :2803-2809
[5]   RESCUE OF A PARALYZED-FLAGELLA MUTANT OF CHLAMYDOMONAS BY TRANSFORMATION [J].
DIENER, DR ;
CURRY, AM ;
JOHNSON, KA ;
WILLIAMS, BD ;
LEFEBVRE, PA ;
KINDLE, KL ;
ROSENBAUM, JL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (15) :5739-5743
[6]  
DUTCHER S, 1995, TRENDS GENET S, V12, P18
[7]   CHLAMYDOMONAS REINHARDI - HETEROZYGOUS DIPLOID STRAINS [J].
EBERSOLD, WT .
SCIENCE, 1967, 157 (3787) :447-&
[8]   ISOLATION AND CHARACTERIZATION OF THE NITRATE REDUCTASE STRUCTURAL GENE OF CHLAMYDOMONAS-REINHARDTII [J].
FERNANDEZ, E ;
SCHNELL, R ;
RANUM, LPW ;
HUSSEY, SC ;
SILFLOW, CD ;
LEFEBVRE, PA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (17) :6449-6453
[9]   COMPONENTS OF A DYNEIN-REGULATORY-COMPLEX ARE LOCATED AT THE JUNCTION BETWEEN THE RADIAL SPOKES AND THE DYNEIN ARMS IN CHLAMYDOMONAS-FLAGELLA [J].
GARDNER, LC ;
OTOOLE, E ;
PERRONE, CA ;
GIDDINGS, T ;
PORTER, ME .
JOURNAL OF CELL BIOLOGY, 1994, 127 (05) :1311-1325
[10]   DYNEIN FAMILY OF MOTOR PROTEINS - PRESENT STATUS AND FUTURE QUESTIONS [J].
GIBBONS, IR .
CELL MOTILITY AND THE CYTOSKELETON, 1995, 32 (02) :136-144