The 26S proteasome in Schistosoma mansoni: bioinformatics analysis, developmental expression, and RNA interference (RNAi) studies.
The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. Here, we used a bioinformatics approach to identify the proteasomal constituents of the parasitic trematode Schistosoma mansoni. A detailed search of the S. mansoni genome database identified a total of 31 putative proteasomal subunits, including 17 subunits of the regulatory (19S) complex and 14 predicted catalytic (20S) subunits. A quantitative real-time RT-PCR analysis of subunit expression levels revealed that the S. mansoni proteasome components are differentially expressed among cercaria, schistosomula, and adult worms. In particular, the data suggest that the proteasome may be downregulated during the early stages of schistosomula development and is subsequently upregulated as the parasite matures to the adult stage. To test for biological relevance, we developed a transfection-based RNA interference method to knockdown the expression of the proteasome subunit, SmRPN11/POH1. Transfection of in vitro transformed S. mansoni schistosomula with specific short-interfering RNAs (siRNAs) diminished SmRPN11/POH1 expression nearly 80%, as determined by quantitative RT-PCR analysis, and also decreased parasite viability 78%, whereas no significant effect could be seen after treatment with the same amount of an irrelevant siRNA. These results indicate that the subunit SmRPN11/POH1 is an essential gene in schistosomes and further suggest an important role for the proteasome in parasite development and survival.
Nabhan
Joseph F
JF
Institute of Parasitology, McGill University, Macdonald Campus, 21,111 Lakeshore Road, Sainte Anne de Bellevue, Que., Canada H9X 3V9.
El-Shehabi
Fouad
F
Patocka
Nicholas
N
Ribeiro
Paula
P
D000818:Animals; D001700:Biomphalaria; D019295:Computational Biology; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008018:Life Cycle Stages / Q000502:physiology; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012550:Schistosoma mansoni / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D016415:Sequence Alignment; D014162:Transfection
D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease