New insights into nuclear import and nucleolar localization of yeast RNA exosome subunits

Author(s)

Neto, Valdir Gomes ; Cepeda, Leidy Paola P. ; Queiroz, Bruno R. S. ; Cantaloube, Sylvain ; Leger-Silvestre, Isabelle ; Mangeat, Thomas ; Albert, Benjamin ; Gadal, Olivier ; Oliveira, Carla C.

• Document type: Journal article
• Source: MOLECULAR BIOLOGY OF THE CELL; v. 36, n. 6, p. 20-pg., 2025-06-01.

Abstract

The RNA exosome is a multiprotein complex essential for RNA maturation and degradation. In budding yeast, a nine-subunit protein core (Exo9) associated with Rrp44 forms a 10-subunit complex (Exo10) in the cytoplasm and, in complex with Rrp6, Exo11 in the nucleus. Depending on its subcellular localization, the exosome interacts with different cofactors and RNA substrates. In the cytoplasm, Exo10 associates with the SKI complex via Ski7, while in the nucleus, Exo11 interacts with the TRAMP complex. Within the nucleolus, the exosome participates in rRNA processing, facilitated by Mtr4-dependent adaptors Utp18 and Nop53. In this article, we have performed a comprehensive study that addresses the targeting mechanism and precise subcellular localization of all members of the Exo11 complex. We observed a high concentration of all Exo11 subunits in the nucleolus and identified the importins Srp1 (alpha) and Kap95 (beta) as responsible for the nuclear import of Exo9 subunits. Notably, Exo9 subunits localization was not significantly disrupted in the simultaneous absence of NLS-containing subunits Rrp6 and Rrp44, suggesting redundant nuclear import pathways for Exo9. Additionally, we show evidence that Ski7 may play a role in the Exo9 retention in the cytoplasm. To explore the exosome subnucleolar localization, we compared Rrp43 with nuclear exosome cofactors and show that it is enriched in the same nucleolar region as Mtr4 and Nop53. In conclusion, our findings provide a detailed characterization of Exo11 distribution, highlight the primary nuclear import mechanisms for Exo9, and reveal the specific localization of the exosome within the granular component of the yeast nucleolus, suggesting a spatial regulation of the RNA-processing pathway. (AU)

• FAPESP's process: 22/16740-2 - Understanding the mechanism of the RNA Exosome subunits nuclear transport in Saccharomyces cerevisiae
• Grantee: Valdir Gomes Neto
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 20/00901-1 - Posttranscriptional control of gene expression: pre-rRNA processing, mRNA degradation, splicing and snoRNP assembly in Saccharomyces cerevisiae
• Grantee: Carla Columbano de Oliveira
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 22/00071-4 - Identification of karyopherins involved in the nuclear import of exosome subunits in Saccharomyces cerevisiae
• Grantee: Valdir Gomes Neto
• Support Opportunities: Scholarships in Brazil - Post-Doctoral