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PROTEOTRANSCRIPTOMIC VIEW ON THE ROLE OF SPIDER SILK GLANDS DURING THE SILK SPINNING PROCESS

Grant number: 16/10430-0
Support Opportunities:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): August 01, 2016
Effective date (End): November 30, 2016
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Organic Chemistry
Principal Investigator:Mario Sergio Palma
Grantee:José Roberto Aparecido dos Santos-Pinto
Supervisor: Gert Lubec
Host Institution: Instituto de Biociências (IB). Universidade Estadual Paulista (UNESP). Campus de Rio Claro. Rio Claro , SP, Brazil
Research place: University of Vienna, Austria  
Associated to the scholarship:13/26451-9 - Bioprospecting and Structural Analysis of the Silk Proteins of Arthropods by a Proteomics Approach Using nanoLC-ESI-CID/ETD System, BP.PD

Abstract

Spider silk proteins are synthesized in the silk-producing glands, where the spidroins are produced, stored and processed into a solid fiber from a crystalline liquid solution. Despite great interest in the spider silk properties, that make this material suitable for biomedical and biotechnological applications, the mechanism of formation and spinning of the silk fibers has not been fully elucidated; and no combination of proteomic and transcriptomic study has been carried out so far in the spider silk-producing glands. Nephila clavipes is an attractive orb-web spider to investigate the spinning process of silk production, given the properties of strength, elasticity and biocompatibility of their silk fibers. Thus, considering that the combination of proteomic and transcriptomic analysis may reveal an extensive repertoire of novel proteins involved in the silk spinning process, and in order to facilitate and enable proteomics in this non-model organism, the current study aims to construct a high quality reference mRNA-derived protein database that could be used to identify tissue specific expression patterns in spider silk glands. Next-generation sequencing has offered a powerful and cost-efficient technique for the generation of transcriptomic datasets in non-model species using diverse platforms such as the Illumina HiSeq, Roche 454, Pacific Biosystems, and Applied Biosystems SOLiD; and mass spectrometry analysis allows large proteome surveys including post-translational modification (PTM) studies and comparative analysis, where quantities of hundreds or thousands of proteins can be compared between various conditions. In the current study, the Illumina HiSeq 2000 platform will be used to generate a N. clavipes spider silk glands transcriptome-based protein database. Moreover, PTMs searches will be performed using Modiro PTM Explorer 1.1 software, which is not available in Brazilian laboratory, to verify PTMs of the spidroin proteins. The transcriptome data generated in this study will provide a comprehensive and valuable genomic resource for future research of the group of spider silk-producing glands, in order to improve our understanding of the overall mechanism of action involved in production, secretion, storage, transport, protection and conformational changes of spidroins during the spinning process, and prey capture; and the results may be relevant for scientists in material Science, biology, biochemistry, and environmental scientists.

News published in Agência FAPESP Newsletter about the scholarship:
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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
APARECIDO DOS SANTOS-PINTO, JOSE ROBERTO; ESTEVES, FRANCIELE GREGO; TORMENA, CLAUDIO FRANCISCO; PEREZ-RIVEROL, AMILCAR; LASA, ALEXIS MUSACCHIO; BUENO, ODAIR CORREA; PALMA, MARIO SERGIO. Proteomic characterization of the fibroin-based silk fibers produced by weaver ant Camponotus textor. JOURNAL OF PROTEOMICS, v. 261, p. 13-pg., . (17/22405-3, 13/26451-9, 17/10373-0, 16/16212-5, 20/10246-0, 16/10430-0, 15/08541-6)
ESTEVES, FRANCIELE GREGO; APARECIDO DOS SANTOS-PINTO, JOSE ROBERTO; FERRO, MILENE; SIALANA, FERNANDO J.; SMIDAK, ROMAN; RARES, LUCACIU CALIN; NUSSBAUMER, THOMAS; RATTEI, THOMAS; BILBAN, MARTIN; BACCI JUNIOR, MAURICIO; et al. Revealing the Venomous Secrets of the Spider's Web. JOURNAL OF PROTEOME RESEARCH, v. 19, n. 8, p. 3044-3059, . (15/14220-8, 17/10373-0, 13/26451-9, 16/10430-0, 16/16212-5)
APARECIDO DOS SANTOS-PINTO, JOSE ROBERTO; ESTEVES, FRANCIELE GREGO; SIALANA, FERNANDO J.; FERRO, MILENE; SMIDAK, ROMAN; RARES, LUCACIU CALIN; NUSSBAUMER, THOMAS; RATTEI, THOMAS; BILBAN, MARTIN; BACCI JUNIOR, MAURICIO; et al. A proteotranscriptomic study of silk-producing glands from the orb-weaving spiders. MOLECULAR OMICS, v. 15, n. 4, p. 256-270, . (16/16212-5, 16/10430-0, 13/26451-9, 17/10373-0, 15/14220-8)

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