Study of the RNA helicase DEAD-box NtDDX35 function

• Grant number: 22/05401-2
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): July 01, 2022
• Effective date (End): June 30, 2024
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Maria Helena de Souza Goldman
• Grantee: Renan Terassi Pinto
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 19/24774-1 - The importance of the SCI1 gene in the floral meristem, its transcriptional regulation and evolutionary conservation, AP.TEM

Abstract

SCI1 is a negative regulator of cell proliferation, characterized by our research group, which affects final pistil size in Nicotiana tabacum and Arabidopsis thaliana. Preliminary results obtained from further explorations of its functionality suggest that SCI1 acts on RNA metabolism, more specifically on transcript splicing, but known interaction sites with RNA in its protein sequence were not identified. This process can occur through interaction with the NtDDX35 protein, which was identified as a partner of SCI1 in a yeast two-hybrid (Y2H) screening, with interaction already confirmed by BiFC, FRET and in vitro pulldown. However, the NtDDX35 protein has not yet been studied in plants and its function is not known. Thus, it is necessary to characterize this possible RNA helicase to understand its influence on the balance between cell proliferation and differentiation in the floral meristem, in addition to other possible functions in plant development. Therefore, from the execution of this project, we intend to answer the following questions: 1) Does the NtDDX35 protein interact directly with RNA?; 2) With which transcripts does this protein interact directly and what are the affinity sites?; 3) Which transcripts are affected in flower meristems/buds, directly or indirectly, as a consequence of changes in the level of NtDDX35 expression in plants? The answers to these questions will advance the understanding of the influence of these possible components of the spliceosome on the balance between cell proliferation and differentiation during the development of the floral meristem. (AU)