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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Nucleic Acids Research; v. 48, n. 10, p. 5511-5526, JUN 4 2020.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 15/13618-8 - Regulando trans-reguladores: investigação da via molecular de PRMT7 como regulador epigenético da virulência em Leishmania
• Beneficiário: Angela Kaysel Cruz
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/19400-1 - Estudo dos efeitos da metilação catalisada por PRMT7 na função e expressão da proteína ligante de RNA Alba20 em Leishmania major
• Beneficiário: Tiago Rodrigues Ferreira
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/50954-3 - Investigação bioquímica da função da enzima PRMT7 na infectividade de parasitos Leishmania spp: varredura de alvos de relevância para Leishmaniose
• Beneficiário: Angela Kaysel Cruz
• Modalidade de apoio: Auxílio à Pesquisa - Regular