: Genes coding for the cuticular proteins (CPs) of Anopheles gambiae will be identified and temporal and spatial patterns of expression of each gene, or group of related genes, will be determined. CPs are among the most abundant multigene families in insects. Over half of the sequences identified to date have a conserved motif, the R&R consensus, which confers the ability to bind chitin. This motif is found in two forms, one associated with proteins from soft, the other with proteins from hard cuticles. CP motifs known from other insects will be used in computer analyses of the Anopheles genome. Once putative CPs are identified, their temporal patterns of expression will be determined with quantitative RT-PCR. Then the spatial location of representative mRNAs will be determined with in situ hybridization on histological sections of animals of different developmental stages. Finding mRNAs in the epidermis underlying a forming cuticle will be taken to indicate that the candidate proteins are destined for secretion into the cuticle. Localization will also provide a correlation of primary protein structure with cuticle type. To learn if putative CPs that lack the R&R consensus also bind chitin, cDNAs for representative proteins will be expressed, their protein products purified and tested for binding on chitin columns. Further information on proteins in cuticle, including those that are not solubilized by strong denaturing agents, will be obtained by using shotgun proteomic methods to identify peptides that can be released by proteases from intact and extracted cuticles. This first comprehensive analysis of the CPs of a single species will clarify the roles that diverse types of CPs play in the structure and function of cuticle, especially as the sequences identified will also be subject to analysis of secondary structure and further structural modeling. Such information that extends from the genome of Anopheles gambiae to the constituents and construction of its cuticle may point the way to new strategies to control this formidable vector.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI055624-04
Application #
7221912
Study Section
Special Emphasis Panel (ZRG1-TMP (99))
Program Officer
Costero, Adriana
Project Start
2004-05-01
Project End
2010-04-30
Budget Start
2007-05-01
Budget End
2010-04-30
Support Year
4
Fiscal Year
2007
Total Cost
$279,144
Indirect Cost
Name
University of Georgia
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Zhou, Yihong; Badgett, Majors J; Orlando, Ron et al. (2018) Proteomics reveals localization of cuticular proteins in Anopheles gambiae. Insect Biochem Mol Biol :
Vannini, Laura; Willis, Judith H (2017) Localization of RR-1 and RR-2 cuticular proteins within the cuticle of Anopheles gambiae. Arthropod Struct Dev 46:13-29
Vannini, Laura; Willis, Judith H (2016) Immunolocalization of cuticular proteins in Johnston's organ and the corneal lens of Anopheles gambiae. Arthropod Struct Dev 45:519-535
Zhou, Yihong; Badgett, Majors J; Bowen, John Hunter et al. (2016) Distribution of cuticular proteins in different structures of adult Anopheles gambiae. Insect Biochem Mol Biol 75:45-57
Vannini, Laura; Bowen, John Hunter; Reed, Tyler W et al. (2015) The CPCFC cuticular protein family: Anatomical and cuticular locations in Anopheles gambiae and distribution throughout Pancrustacea. Insect Biochem Mol Biol 65:57-67
Ioannidou, Zoi S; Theodoropoulou, Margarita C; Papandreou, Nikos C et al. (2014) CutProtFam-Pred: detection and classification of putative structural cuticular proteins from sequence alone, based on profile hidden Markov models. Insect Biochem Mol Biol 52:51-9
Vannini, Laura; Reed, Tyler W; Willis, Judith H (2014) Temporal and spatial expression of cuticular proteins of Anopheles gambiae implicated in insecticide resistance or differentiation of M/S incipient species. Parasit Vectors 7:24
Vannini, Laura; Augustine Dunn, W; Reed, Tyler W et al. (2014) Changes in transcript abundance for cuticular proteins and other genes three hours after a blood meal in Anopheles gambiae. Insect Biochem Mol Biol 44:33-43
Chipman, Ariel D; Ferrier, David E K; Brena, Carlo et al. (2014) The first myriapod genome sequence reveals conservative arthropod gene content and genome organisation in the centipede Strigamia maritima. PLoS Biol 12:e1002005
Werren, John H; Richards, Stephen; Desjardins, Christopher A et al. (2010) Functional and evolutionary insights from the genomes of three parasitoid Nasonia species. Science 327:343-8

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