Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Débora N. Santos, Pedro H.N. Aguiar, Francisco P. Lobo, Marina M. Mourão, José H.M. Tambor, Analina F. Valadão, Adlane Vilas-Boas, Francisco G. Nobrega, Philip T. LoVerde, Andréa M. Macedo, Sérgio D.J. Pena, Carlos R. Machado, Glória R. Franco

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1- yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxΔ24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation.

Original languageEnglish (US)
Pages (from-to)440-449
Number of pages10
JournalExperimental Parasitology
Volume116
Issue number4
DOIs
StatePublished - Aug 1 2007

Keywords

  • HRT1
  • RING box
  • SCF complex
  • Schistosoma mansoni
  • SmRbx
  • Trematode
  • Ubiquitination

ASJC Scopus subject areas

  • Parasitology
  • Immunology
  • Infectious Diseases

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  • Cite this

    Santos, D. N., Aguiar, P. H. N., Lobo, F. P., Mourão, M. M., Tambor, J. H. M., Valadão, A. F., Vilas-Boas, A., Nobrega, F. G., LoVerde, P. T., Macedo, A. M., Pena, S. D. J., Machado, C. R., & Franco, G. R. (2007). Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination. Experimental Parasitology, 116(4), 440-449. https://doi.org/10.1016/j.exppara.2007.02.012