Study of SCI1 and its protein-protein interactions that result in cell cycle inhibition

• Grant number: 12/50562-2
• Support Opportunities: Regular Research Grants
• Start date: November 01, 2012
• End date: April 30, 2015
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Maria Helena de Souza Goldman
• Grantee: Maria Helena de Souza Goldman
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

In our laboratory it was identified the SCH gene (Stigma/style Cell cyde Inhibitor 1), capable of controlling cell proliferation on the upper part of the pistil (DePaoli et ai., 2011), probably through the inhibrtron of the cyclin-CDK (Cyclin Dependent protein Kinase) complex. To exert its function, the protein SCH has to interact with other proteins. In a previous project (process no. 10/50213-2), some candidates for protein interaction were identified, like the CDKG;2 and two proteins of the 14-3-3 family. Therefore, the objectives of this project are: 1) To identify/confirm the cyclin(s) that interacts with SC/1; 2) To identify partners/substrates of the cyclin dependent kinase (CDKG;2), by the screening of a stigma/style library in the two hybrid vector; 3) To study the hypothesis of SCH regulating cell division through the inhibition of CDKG.2. a. To test the interaction of the proteins cyclin L and other cyclins (cyclin B1 and/or cyclin-related) with CDKG;2 by two hybrid and/or BiFC (Bimolecular Fluorescence Complementation). b. In the case a cyclin capable of interacting with CDKG;2 is identified in the directed experiments (item 3.a) or in the screening, to test CDKG;2 activity on histones and/or another possible substrate identified on the screening (item 2); c. If a and b are positive, to test the inhibition of CDKG;2 activity by SCI1; 4) To test the hypothesis of tissue-specificity on the expression of SCH interaction partners; 5) To analyze the subcellular localization of the CDKG;2,14-3-3A and 14-3-3D proteins; 6) To perform two hybrid and/or BiFC experiments to test the formation of homo and heterodimers between the 2 proteins of the 14-3-3 family and their interactions with SCH; 7) To characterize the phosphorylation of SCH protein, by the comparison of phosphorylated residues in SCH extracted from leaves and pistils of 35Sprom::SCI1-GFP transgenic plants; 8) To confirm SCI1 interaction with the new candidates identified by pull-down or two hybrid screening performed. (AU)

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)

STRINI, EDWARD J.; BERTOLINO, LIGIA T.; SAN MARTIN, JUCA A. B.; SOUZA, HEBREIA A. O.; PESSOTTI, FRANCINE; PINOTI, VITOR F.; FERREIRA, PEDRO B.; DE PAOLI, HENRIQUE C.; LUBINI, GREICE; DEL-BEM, LUIZ-EDUARDO; et al. Stigma/Style Cell-Cycle Inhibitor 1, a Regulator of Cell Proliferation, Interacts With a Specific 14-3-3 Protein and Is Degraded During Cell Division. FRONTIERS IN PLANT SCIENCE, v. 13, p. 14-pg., 2022-04-04. (19/24774-1, 16/20486-3, 12/50562-2, 11/51844-9)