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Molecular cloning of a functional allatostatin gut/brain receptor and an allatostatin preprohormone from the silkworm Bombyx mori.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2001 Dec 14; Vol. 276 (50), pp. 47052-60. Date of Electronic Publication: 2001 Oct 04. - Publication Year :
- 2001
-
Abstract
- The cockroach-type or A-type allatostatins are inhibitory insect neuropeptides with the C-terminal sequence Tyr/Phe-X-Phe-Gly-Leu-NH(2). Here, we have cloned an A-type allatostatin receptor from the silkworm Bombyx mori (BAR). BAR is 361 amino acid residues long, has seven transmembrane domains, shows 60% amino acid residue identity with the first Drosophila allatostatin receptor (DAR-1), and 48% identity with the second Drosophila allatostatin receptor (DAR-2). The BAR gene has two introns and three exons. These two introns coincide with and have the same intron phasing as two introns in the DAR-1 and DAR-2 genes, showing that the three receptors are not only structurally but also evolutionarily related. Furthermore, we have cloned a Bombyx allatostatin preprohormone that contains eight different A-type allatostatins. Chinese hamster ovary cells permanently transfected with BAR DNA react on the addition of 4 x 10(-9)M Bombyx A-type allatostatins with a second messenger cascade (measured as bioluminescence), showing that BAR is a functional A-type allatostatin receptor. Southern blots suggest that Bombyx has at least one other BAR-related gene in addition to the BAR gene described in this paper. Northern blots and quantitative reverse transcriptase-polymerase chain reaction of different larval tissues show that BAR mRNA is mainly expressed in the gut and to a much lesser extent in the brain. To our knowledge, this is the first report on the molecular cloning and functional expression of an insect gut/brain peptide hormone receptor.
- Subjects :
- Amino Acid Sequence
Animals
Blotting, Northern
Blotting, Southern
Bombyx
CHO Cells
Cloning, Molecular
Cricetinae
DNA, Complementary metabolism
Dose-Response Relationship, Drug
Drosophila
Evolution, Molecular
Exons
Introns
Kinetics
Molecular Sequence Data
Phylogeny
RNA, Messenger metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
Signal Transduction
Tissue Distribution
Transfection
Brain metabolism
Digestive System metabolism
Drosophila Proteins
Hormones genetics
Insect Proteins
Neuropeptides
Protein Precursors genetics
Receptors, Cell Surface chemistry
Receptors, Cell Surface genetics
Receptors, Cell Surface metabolism
Receptors, G-Protein-Coupled
Receptors, Neuropeptide
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9258
- Volume :
- 276
- Issue :
- 50
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 11590150
- Full Text :
- https://doi.org/10.1074/jbc.M106675200