Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit

Submitted by Manoel Barata ([email protected]) on 2018-09-14T12:28:06Z No. of bitstreams: 1 JournalBiolChemistryzb4853.pdf: 1990964 bytes, checksum: bb68ee6b122af42a80fca5054d5c4a3d (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2018-09-14T19:47:44Z (GMT) No. of bitstreams: 1 JournalBiolChemistryzb4853.pdf: 1990964 bytes, checksum: bb68ee6b122af42a80fca5054d5c4a3d (MD5)Made available in DSpace on 2018-09-14T19:47:44Z (GMT). No. of bitstreams: 1 JournalBiolChemistryzb4853.pdf: 1990964 bytes, checksum: bb68ee6b122af42a80fca5054d5c4a3d (MD5) Previous issue date: 2012Centro Nacional de Pesquisa em Energia e Materiais. Laboratório Nacional de Biociências. Campinas, SP, Brasil. / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.Centro Nacional de Pesquisa em Energia e Materiais. Laboratório Nacional de Biociências. Campinas, SP, Brasil.Universidade Estadual de Campinas. Centro de Biologia Molecular e Engenharia Genética, Campinas, SP, Brasil.Centro Nacional de Pesquisa em Energia e Materiais. Laboratório Nacional de Biociências. Campinas, SP, Brasil. / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.Universidade Estadual de Campinas. Centro de Biologia Molecular e Engenharia Genética, Campinas, SP, Brasil. / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.PP2A is the main serine/threonine-specific phosphatase in animal cells. The active phosphatase has been described as a holoenzyme consisting of a catalytic, a scaffolding, and a variable regulatory subunit, all encoded by multiple genes, allowing for the assembly of more than 70 different holoenzymes. The catalytic subunit can also interact with α4, TIPRL (TIP41, TOR signaling pathway regulator-like), the methyl-transferase LCMT-1, and the methyl-esterase PME-1. Here, we report that the gene encoding the catalytic subunit PP2Acα can generate two mRNA types, the standard mRNA and a shorter isoform, lacking exon 5, which we termed PP2Acα2. Higher levels of the PP2Acα2 mRNA, equivalent to the level of the longer PP2Acα mRNA, were detected in peripheral blood mononuclear cells that were left to rest for 24 h. After this time, the peripheral blood mononuclear cells are still viable and the PP2Acα2 mRNA decreases soon after they are transferred to culture medium, showing that generation of the shorter isoform depends on the incubation conditions. FLAG-tagged PP2Acα2 expressed in HEK293 is catalytically inactive. It displays a specific interaction profile with enhanced binding to the α4 regulatory subunit, but no binding to the scaffolding subunit and PME-1. Consistently, α4 out-competes PME-1 and LCMT-1 for binding to both PP2Acα isoforms in pulldown assays. Together with molecular modeling studies, this suggests that all three regulators share a common binding surface on the catalytic subunit. Our findings add important new insights into the complex mechanisms of PP2A regulation

Cyclic lipopeptide signature as fingerprinting for the screening of halotolerant bacillus strains towards microbial enhanced oil recovery

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIORCyclic lipopeptides (CLPs) are non-ribosomal biosurfactants produced by Bacillus species that exhibit outstanding interfacial activity. The synthesis of CLPs is under genetic and environmental influence, and representatives from different families are gen10211791190CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIORCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIORsem informaçãosem informaçãoWe thank André Saraiva L M Antunes for proof reading the manuscript. We acknowledge PETROBRAS for the financial support (Cooperation agreement: 0050.0079828.12.9) and authorization to publish this work and Coleção de Culturas do Gênero Bacillus e Gênero