PRMT7 regulates RNA-binding capacity and protein stability in Leishmania parasites

Author(s)

Ferreira, Tiago R. ^{[1, 2]} ; Dowle, Adam A. ^[3] ; Parry, Ewan ^[1] ; Alves-Ferreira, Eliza V. C. ^[1] ; Hogg, Karen ^[4] ; Kolokousi, Foteini ^[1] ; Larson, Tony R. ^[3] ; Plevin, Michael J. ^[1] ; Cruz, Angela K. ^[5] ; Walrad, Pegine B. ^[1]

Affiliation

^[1] Univ York, York Biomed Res Inst, Dept Biol, York, N Yorkshire - England

^[2] NIAID, NIH, 9000 Rockville Pike, Bethesda, MD 20892 - USA

^[3] Univ York, Dept Biol, Metabol & Prote Lab, Biosci Technol Facil, York, N Yorkshire - England

^[4] Univ York, Dept Biol, Imaging & Cytometry Lab, Biosci Technol Facil, York, N Yorkshire - England

^[5] Univ Sao Paulo, Ribeirao Preto Med Sch, Cell & Mol Biol Dept, Ribeirao Preto - Brazil

• Document type: Journal article
• Source: Nucleic Acids Research; v. 48, n. 10, p. 5511-5526, JUN 4 2020.
• Web of Science Citations: 0

Abstract

RNA binding proteins (RBPs) are the primary gene regulators in kinetoplastids as transcriptional control is nearly absent, making Leishmania an exceptional model for investigating methylation of non-histone substrates. Arginine methylation is an evolutionarily conserved protein modification catalyzed by Protein aRginine Methyl Transferases (PRMTs). The chromatin modifier PRMT7 is the only Type III PRMT found in higher eukaryotes and a restricted number of unicellular eukaryotes. In Leishmania major, PRMT7 is a cytoplasmic protein implicit in pathogenesis with unknown substrates. Using comparative methyl-SILAC proteomics for the first time in protozoa, we identified 40 putative targets, including 17 RBPs hypomethylated upon PRMT7 knockout. PRMT7 can modify Alba3 and RBP16 trans-regulators (mammalian RPP25 and YBX2 homologs, respectively) as direct substrates in vitro. The absence of PRMT7 levels in vivo selectively reduces Alba3 mRNA-binding capacity to specific target transcripts and can impact the relative stability of RBP16 in the cytoplasm. RNA immunoprecipitation analyses demonstrate PRMT7-dependent methylation promotes Alba3 association with select target transcripts and thus indirectly stabilizes mRNA of a known virulence factor, delta-amastin surface antigen. These results highlight a novel role for PRMT7-mediated arginine methylation of RBP substrates, suggesting a regulatory pathway controlling gene expression and virulence in Leishmania. This work introduces Leishmania PRMTs as epigenetic regulators of mRNA metabolism with mechanistic insight into the functional manipulation of RBPs by methylation. (AU)

• FAPESP's process: 15/13618-8 - Regulating the trans-regulators: investigating the PRMT7 molecular pathway as an epigenetic regulator of Leishmania virulence
• Grantee: Angela Kaysel Cruz
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/19400-1 - Study of the effects of PRMT7-catalyzed methylation on the function and expression of the RNA-binding protein Alba20 in Leishmania major
• Grantee: Tiago Rodrigues Ferreira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50954-3 - Biochemical investigation of the enzyme PRMT7 function in Leishmania spp. parasite infectivity: screening targets for Leishmaniasis relevance
• Grantee: Angela Kaysel Cruz
• Support Opportunities: Regular Research Grants