Molecular characterization and distribution of CYCLE protein from Athalia rosae.
cDNA encoding CYCLE (CYC) from the coleseed sawfly, Athalia rosae (Hymenoptera, Symphyta), was amplified by PCR. This is a first determination of hymenopteran CYC structure. ArCYC had an overall identity of 66% with CYC of Anopheles gambiae and ca. 60% of Drosophila melanogaster. Structural investigation revealed that ArCYC contained characteristic motifs of: bHLH, PAS A, PAS B, PAC and BCTR. Detailed analysis indicated high conservation of these regions among insects. Northern blot analysis showed that the mRNA of ca. 3 kb was transcribed both in the head and in the rest of the body. Southern blot analysis suggested the presence of a single copy of the gene in the genome. Western blot indicates that the quantity of CYC protein does not fluctuate under LD 12:12 in either the head or the rest of the body. Immunocytochemical examination revealed CYC-like antigen in the pars intercerebralis, dorsolateral protocerebrum, dorsal optic tract, tritocerebrum of the brain and the subesophageal ganglion.
Bembenek
Jadwiga
J
Graduate School of Science and Technology, Kobe University, Kobe 657 8501, Japan. jadwigabembenek@yahoo.com
Itokawa
Kentaro
K
Hiragaki
Susumu
S
Qi-Miao Shao
Tufail
Muhammad
M
Takeda
Makio
M
D056930:ARNTL Transcription Factors; D000595:Amino Acid Sequence; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000737:chemistry; D001683:Biological Clocks / Q000502:physiology*; D002490:Central Nervous System / Q000378:metabolism; D002940:Circadian Rhythm / Q000502:physiology*; D029721:Drosophila Proteins / Q000737:chemistry; D006927:Hymenoptera / Q000378:metabolism* / Q000502:physiology; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D056930:ARNTL Transcription Factors; D051792:Basic Helix-Loop-Helix Proteins; D029721:Drosophila Proteins; D019476:Insect Proteins; C112805:cyc protein, Drosophila