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The Chlamydomonas Dhc1 gene encodes a dynein heavy chain subunit required for assembly of the I1 inner arm complex.
- Source :
-
Molecular biology of the cell [Mol Biol Cell] 1997 Apr; Vol. 8 (4), pp. 607-20. - Publication Year :
- 1997
-
Abstract
- Multiple members of the dynein heavy chain (Dhc) gene family have been recovered in several organisms, but the relationships between these sequences and the Dhc isoforms that they encode are largely unknown. To identify Dhc loci and determine the specific functions of the individual Dhc isoforms, we have screened a collection of motility mutants generated by insertional mutagenesis in Chlamydomonas. In this report, we characterize one strain, pf9-3, in which the insertion event was accompanied by a deletion of approximately 13 kb of genomic DNA within the transcription unit of the Dhc1 gene. Northern blot analysis confirms that pf9-3 is a null mutation. Biochemical and structural studies of isolated axonemes demonstrate that the pf9-3 mutant fails to assemble the I1 inner arm complex, a two-headed dynein isoform composed of two Dhcs (1 alpha and 1 beta) and three intermediate chains. To determine if the Dhc1 gene product corresponds to one of the Dhcs of the I1 complex, antibodies were generated against a Dhc1-specific peptide sequence. Immunoblot analysis reveals that the Dhc1 gene encodes the 1 alpha Dhc subunit. These studies thus, identify the first inner arm Dhc locus to be described in any organism and further demonstrate that the 1 alpha Dhc subunit plays an essential role in the assembly of the I1 inner arm complex.
- Subjects :
- Amino Acid Sequence
Animals
Binding Sites
Cell Movement genetics
Chlamydomonas physiology
Chromosome Mapping
DNA Transposable Elements
Dyneins isolation & purification
Gene Deletion
Isoenzymes
Molecular Sequence Data
Multigene Family
Mutation
Phenotype
Transcription, Genetic
Chlamydomonas genetics
Dyneins genetics
Dyneins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1059-1524
- Volume :
- 8
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Molecular biology of the cell
- Publication Type :
- Academic Journal
- Accession number :
- 9247642
- Full Text :
- https://doi.org/10.1091/mbc.8.4.607