Functional study of the components of the Saccharomyces cerevisiae exosome and the spliceosome, and their role in posttranscriptional control of gene expression

• Grant number: 15/06477-9
• Support Opportunities: Research Projects - Thematic Grants
• Duration: January 01, 2016 - December 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Carla Columbano de Oliveira
• Grantee: Carla Columbano de Oliveira
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated grant(s): 17/17706-4 - Multi-user equipment approved in grant 15/06477-9: ultra-centrifuge and rotors, AP.EMU
• Associated scholarship(s): 20/15106-2 - Study of the exosome recruitment mechanism in the initial stages of ribosomal biogenesis, BP.IC; 20/03315-6 - Construction and characterization of thermosensitive mutants of the general splicing-factor Cwc24 in Saccharomyces cerevisiae, BP.IC; 18/17562-5 - Characterization of neuronal proteins that interact with the EXOSC8 (Rrp43) subunit of the exosome and regulate its activity, BP.DD; 17/17777-9 - Study of the celular transport mechanism of the exosome's subunits in Saccharomyces cerevisiae, BP.MS

Abstract

In eukaryotes, RNAs undergo many processing steps for becoming functional; these steps require specific protein-protein and protein-RNA interactions that are essential for the control of gene expression. We have characterized the function of various yeast proteins, involved in different steps of RNA processing. The research in our group has focused mainly in the study of the exosome complex, present in organisms ranging from archaea to mammals, in which it is involved in processing and degradation of all classes of RNAs. We have been studying the function of the exosome in the yeast Saccharomyces cerevisiae and the structure of the archaeal and yeast complexes. In this project, we intend to continue studying the exosome, mainly analyzing the subcelular localization of this complex in different mutants. Nop53 has been characterized in our laboratory as a factor involved in rRNA processing. Nop53 interacts with the exosome subunit Rrp6, and activates this complex during processing in the nucleolus. In this project, we intend to continue those studies and determine the mechanism by which Nop53 activates the exosome. Among the proteins studied by our group, Cwc24 was shown to be a general splicing factor interacting with Brr2 and Prp19, although it is mainly important for splicing of precursor RNAs containing non-consensus branch site sequences. In this project, we intend to continue the study of Cwc24 function for the determination of its role in splicing. This project, therefore, proposes the study of various aspects of RNA processing, which involve different enzymatic complexes, interconnected by protein interactions. (AU)

Scientific publications

(6)

(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)

BAGATELLI, FELIPE F. M.; DE LUNA VITORINO, FRANCISCA N.; DA CUNHA, JULIA P. C.; OLIVEIRA, CARLA C.. The ribosome assembly factor Nop53 has a structural role in the formation of nuclear pre-60S intermediates, affecting late maturation events. Nucleic Acids Research, v. 49, n. 12, p. 7053-7074, JUL 9 2021. (15/05776-2, 15/06477-9, 13/07467-1)

CEPEDA, LEIDY PAOLA P.; BAGATELLI, FELIPE F. M.; SANTOS, RENATA M.; SANTOS, MARLON D. M.; NOGUEIRA, FABIO C. S.; OLIVEIRA, CARLA C.. The ribosome assembly factor Nop53 controls association of the RNA exosome with pre-60S particles in yeast. Journal of Biological Chemistry, v. 294, n. 50, p. 19365+, DEC 13 2019. (15/06477-9, 15/05776-2)

GONZALES-ZUBIATE, FERNANDO A.; OKUDA, ELLEN K.; DA CUNHA, JULIA P. C.; OLIVEIRA, CARLA COLUMBANO. Identification of karyopherins involved in the nuclear import of RNA exosome subunit Rrp6 in Saccharomyces cerevisiae. Journal of Biological Chemistry, v. 292, n. 29, p. 12267-12284, JUL 21 2017. (12/50196-6, 10/51842-3, 15/06477-9)

OKUDA, ELLEN K.; GONZALES-ZUBIATE, FERNANDO A.; GADAL, OLIVIER; OLIVEIRA, CARLA C.. Nucleolar localization of the yeast RNA exosome subunit Rrp44 hints at early pre-rRNA processing as its main function. Journal of Biological Chemistry, v. 295, n. 32, p. 11195-11213, AUG 7 2020. (17/17777-9, 15/06477-9, 18/19451-6)

CARVALHO, FELIPE A.; BARROS, MOISES R. A.; GIROTTO, THIERRY P. F.; PERONA, M. GRISELDA; OLIVEIRA, CARLA C.. Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, n. 165, NOV 2020. (15/06477-9)

CARVALHO, FELIPE A.; BARROS, MOISES R. A.; GIROTTO, THIERRY P. F.; PERONA, M. GRISELDA; OLIVEIRA, CARLA C.. Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, v. N/A, n. 165, p. 9-pg., 2020-11-01. (15/06477-9)