Leishmania major phosphoglycerate kinase transcript and protein stability contributes to differences in isoform expression levels

Autor(es)

Azevedo, Aline ^[1] ; Toledo, Juliano S. ^{[1, 2]} ; Defina, Tania ^[1] ; Pedrosa, Andre L. ^[3] ; Cruz, Angela K. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Biol Celular & Mol & Bioagentes Patogen, BR-14049900 Ribeirao Preto - Brazil

^[2] Univ Fed Minas Gerais, Fac Farm, Dept Anal Clin & Toxicol, Belo Horizonte, MG - Brazil

^[3] Univ Fed Triangulo Mineiro, Dept Bioquim Farmacol & Fisiol, Uberaba, MG - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Experimental Parasitology; v. 159, p. 222-226, DEC 2015.
• Citações Web of Science: 1

Resumo

Leishmania contains two phosphoglycerate kinase (PGK) genes, PGKB and PGKC, which code for the cytosolic and glycosomal isoforms of the enzyme, respectively. Although differences in PGKB and PGKC transcript and protein levels and isoform activities have been well documented, the mechanisms of control of both transcript and protein abundance have not been described to date. To better understand the regulation of Leishmania PGK expression, we investigated the stabilities of both PGK transcripts using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) in combination with transcription and trans-splicing inhibitors. Cells were treated with sinefungin and actinomycin D, and RNA decay kinetics were assessed. In addition, immunoblotting and protein synthesis inhibition by cycloheximide were employed to evaluate protein steady states and degradation. We observed increased stabilities of both PGKB mRNA and protein compared with the glycosomal isoform (PGKC). Our results indicate that both post-transcriptional and post-translational events contribute to the distinct expression levels of the PGKB and PGKC isoforms in Leishmania major. (C) 2015 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 04/00859-2 - Investigacao sobre elementos envolvidos no controle de expressao genica em leishmania: os genes das fosfoglicerato-quinases como modelos.
• Beneficiário: Aline de Azevedo
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 99/12403-3 - Do genoma à biologia de Leishmania: uma abordagem compreensiva
• Beneficiário: Angela Kaysel Cruz
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 06/50323-7 - Estudo do controle da expressão gênica e plasticidade genética em Leishmania
• Beneficiário: Angela Kaysel Cruz
• Modalidade de apoio: Auxílio à Pesquisa - Temático