Isolation of dynein heavy chain cDNAs from trout testis which predict an extensive carboxyl-terminal alpha-helical coiled-coil domain.
A Mg2+-dependent ATPase activity has been purified from trout sperm axonemes which has properties characteristic of a dynein ATPase. A polyclonal antiserum prepared against the dynein heavy chains has been used to isolate dynein heavy chain (DYHC) cDNAs from a trout testis lambda gt11 cDNA expression library. beta-galactosidase fusion proteins produced in lambda gt11 by these trout cDNAs cross-reacted with a heterologous anti-sea urchin dynein antiserum. Northern blot analyses demonstrated that the RNA transcripts detected have sizes (7.5 - 12 kb) consistent with those expected for the dynein heavy chains. All the DYHC cDNAs encode portions of a highly unusual DNA coding sequence comprised of 21 bp direct repeats. The predicted open reading frame of this repeat is Ile/Leu-His-Val-Ile-Gln-Tyr-Ser and is characteristic of an extensive alpha-helical coiled-coil domain. The presence of an in-frame translation termination codon indicates that this domain is located at the carboxyl-terminus of the DYHC. Southern blot analyses demonstrated a low, if not single, copy number for this gene and conservation of this domain in other vertebrates. DYHC transcripts reach their highest level in testis, but are also abundant in brain tissue.
Garber
A T
AT
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Retief
J D
JD
Dixon
G H
GH
D000251:Adenosine Triphosphatases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004398:Dyneins / Q000235:genetics*; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011487:Protein Conformation; D012091:Repetitive Sequences, Nucleic Acid; D013737:Testis / Q000201:enzymology; D014158:Transcription, Genetic; D014337:Trout
D004247:DNA; D000251:Adenosine Triphosphatases; D004398:Dyneins