| Sequence of a cDNA and expression of the gene encoding a putative epidermal chitin synthase of Manduca sexta. | |
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MedLine Citation:
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PMID: 12530217 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Glycosyltransferases are enzymes that synthesize oligosaccharides, polysaccharides and glycoconjugates. One type of glycosyltransferase is chitin synthase, a very important enzyme in biology, which is utilized by insects, fungi, and other invertebrates to produce chitin, a polysaccharide of beta-1,4-linked N-acetylglucosamine. Chitin is an important component of the insect's exoskeletal cuticle and gut lining. To identify and characterize a chitin synthase gene of the tobacco hornworm, Manduca sexta, degenerate primers were designed from two highly conserved regions in fungal and nematode chitin synthase protein sequences and then used to amplify a similar region from Manduca cDNA. A full-length cDNA of 5152 nucleotides was assembled for the putative Manduca chitin synthase gene, MsCHS1, and sequencing of genomic DNA verified the contiguity of the sequence. The MsCHS1 cDNA has an ORF of 4692 nucleotides that encodes a transmembrane protein of 1564 amino acid residues with a mass of approximately 179 kDa (GenBank no. AY062175). It is most similar, over its entire length of protein sequence, to putative chitin synthases from other insects and nematodes, with 68% identity to enzymes from both the blow fly, Lucilia cuprina, and the fruit fly, Drosophila melanogaster. The similarity with fungal chitin synthases is restricted to the putative catalytic domain, and the MsCHS1 protein has, at equivalent positions, several amino acids that are essential for activity as revealed by mutagenesis of the fungal enzymes. A 5.3-kb transcript of MsCHS1 was identified by northern blot hybridization of RNA from larval epidermis, suggesting that the enzyme functions to make chitin deposited in the cuticle. Further examination by RT-PCR showed that MsCHS1 expression is regulated in the epidermis, with the amount of transcript increasing during phases of cuticle deposition. |
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Authors:
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Yu-Cheng Zhu; Charles A Specht; Neal T Dittmer; Subbaratnam Muthukrishnan; Michael R Kanost; Karl J Kramer |
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Publication Detail:
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Type: Comparative Study; 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. |
Journal Detail:
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Title: Insect biochemistry and molecular biology Volume: 32 ISSN: 0965-1748 ISO Abbreviation: Insect Biochem. Mol. Biol. Publication Date: 2002 Nov |
Date Detail:
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Created Date: 2003-01-17 Completed Date: 2003-04-30 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 9207282 Medline TA: Insect Biochem Mol Biol Country: England |
Other Details:
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Languages: eng Pagination: 1497-506 Citation Subset: IM |
Affiliation:
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Grain Marketing and Production Research Center, ARS-USDA, 1515 College Avenue, Manhattan, KS 66502, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Cell Membrane / enzymology, ultrastructure Chitin Synthase / genetics* Cloning, Molecular DNA, Complementary Drosophila Epidermis / enzymology Genes, Insect* Manduca / enzymology, genetics* Molecular Sequence Data Protein Conformation Restriction Mapping Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Amino Acid |
| Grant Support | |
ID/Acronym/Agency:
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GM 31318/GM/NIGMS NIH HHS; R01 A125780//PHS HHS |
| Chemical | |
Reg. No./Substance:
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0/DNA, Complementary; EC 2.4.1.16/Chitin Synthase |
| Comments/Corrections | |
Erratum In:
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Insect Biochem Mol Biol. 2003 Feb;33(2):277 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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