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PMID: 9247642 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The Chlamydomonas Dhc1 gene encodes a dynein heavy chain subunit required for assembly of the I1 inner arm complex.

Molecular biology of the cell ·Vol. 8 ·No. 4 ·1997-04-00 ·Pages 607-20

Myster SH, Knott JA, O'Toole E, Porter ME

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.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Cell Movement/genetics Chlamydomonas/genetics,physiology Chromosome Mapping DNA Transposable Elements Dyneins/genetics,isolation & purification,metabolism Gene Deletion Isoenzymes Molecular Sequence Data Multigene Family Mutation Phenotype Transcription, Genetic
Chemicals
DNA Transposable Elements Isoenzymes Dyneins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Myster S H
Department of Cell Biology and Neuroanatomy, University of Minnesota Medical School, Minneapolis 55455, USA.
Knott J A
O'Toole E
Porter M E
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.8.4.607
Published
1997-04-00
Pages
607-20
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC276113
Subset
IM
Grants
NCRR NIH HHS · RR00592 · United States
External Links
PubMed source DOI: 10.1091/mbc.8.4.607 PMC Full Text View journal details
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