Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas.
Flagellar motility is generated by the activity of multiple dynein motors, but the specific role of each dynein heavy chain (Dhc) is largely unknown, and the mechanism by which the different Dhcs are targeted to their unique locations is also poorly understood. We report here the complete nucleotide sequence of the Chlamydomonas Dhc1 gene and the corresponding deduced amino acid sequence of the 1alpha Dhc of the I1 inner dynein arm. The 1alpha Dhc is similar to other axonemal Dhcs, but two additional phosphate binding motifs (P-loops) have been identified in the NH(2)- and COOH-terminal regions. Because mutations in Dhc1 result in motility defects and loss of the I1 inner arm, a series of Dhc1 transgenes were used to rescue the mutant phenotypes. Motile cotransformants that express either full-length or truncated 1alpha Dhcs were recovered. The truncated 1alpha Dhc fragments lacked the dynein motor domain, but still assembled with the 1beta Dhc and other I1 subunits into partially functional complexes at the correct axoneme location. Analysis of the transformants has identified the site of the 1alpha motor domain in the I1 structure and further revealed the role of the 1alpha Dhc in flagellar motility and phototactic behavior.
Myster
S H
SH
Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Knott
J A
JA
Wysocki
K M
KM
O'Toole
E
E
Porter
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D002696:Chlamydomonas / Q000502:physiology*; D003360:Cosmids / Q000235:genetics; D004398:Dyneins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005407:Flagella / Q000737:chemistry / Q000502:physiology*; D005816:Genetic Complementation Test; D020409:Molecular Motor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009068:Movement; D009154:Mutation; D010641:Phenotype; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic; D019076:Transgenes / Q000235:genetics
D020409:Molecular Motor Proteins; D012333:RNA, Messenger; D004398:Dyneins