Investigation on the 19S ATPase proteasome subunits (Rpt1-6) conservation and their differential gene expression in Schistosoma mansoni.
The ubiquitin-proteasome system is responsible for degradation of the majority of intracellular proteins in eukaryotic cells. The 26S proteasome proteolytic complex is composed of a 20S core particle responsible for protein degradation and the 19S lid which plays a role in the recognition of polyubiquitinated substrates. The 19S regulatory particle (Rps) is composed of ATPase (Rpt) and non-ATPase (Rpn) subunits. In this study, we analyzed the expression profile of 19S Rpt subunits in the larvae and adult stage of the Schistosoma mansoni life cycle. Conventional reverse transcriptase polymerase chain reaction (RT-PCR) revealed that the majority of the 19S Rpt subunits amplified at the expected molecular masses for various investigated stages. In addition, SmRpt1, SmRpt2, and SmRpt6 transcript levels were increased in 3 h-cultured schistosomula and reasonably maintained until 5 h in culture, as revealed by qRT-PCR. Phylogenetic analysis of 19S Rpt subunits showed high structural conservation in comparison to other Rpt orthologues. The mRNA expression profile of 19S Rpt subunits did not correlate with 26S proteasome proteolytic activity as judged by a (14)C-casein-degrading assay, in the early cultured schistosomula. Taken together, these results revealed a differential expression profile for 19S Rpt subunits whose transcript levels could not be directly associated to 26S proteasome activity.
Centro de Ciências Agrárias-CCA/UFES, Departamento de Zootecnia, Alto Universitário, s/n Centro, 29500-000, Alegre, Espírito Santo, Brazil.
Olavo S
OS
Pereira-Júnior
Roberta Verciano
RV
Pereira
Camila S
CS
Silva
William
W
Castro-Borges
Renata Guerra
RG
Sá
Fernanda J
FJ
Cabral
Sérgio H
SH
Silva
Cláudia S
CS
Soares
Enyara R
ER
Morais
Erika B C
EB
Moreira
Lizandra G
LG
Magalhães
Fabiana M
FM
de Paula
Vanderlei
V
Rodrigues
D000251:Adenosine Triphosphatases / Q000235:genetics; D000818:Animals; D017124:Conserved Sequence; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007814:Larva / Q000201:enzymology / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010802:Phylogeny; D046988:Proteasome Endopeptidase Complex / Q000235:genetics*; D021122:Protein Subunits / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics*
D021122:Protein Subunits; D012333:RNA, Messenger; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases