Y-box binding protein from Schistosoma mansoni: Interaction with DNA and RNA

A. F. Valadão, M. R. Fantappie, P. T. LoVerde, S. D.J. Pena, F. D. Rumjanek, G. R. Franco

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A Schistosoma mansoni homologue of the human Y-box binding protein (SMYB1), as well as truncated proteins containing its N-terminal Cold Shock Domain (CSD) or its C-terminal domain (TAIL) were cloned into the p-MAL-c2 expression vector and produced in Escherichia coli. In order to characterize the interactions of these proteins to an inverted CCAAT motif present in a number of gene promoters, their binding to DNA was measured by Electrophoretic Mobility Shift Assays. SMYB1 bound to single- and double-stranded DNA containing the CCAAT motif and could bind also to RNA. The truncated CSD and TAIL domain proteins bound to dsDNA and RNA, but exhibited distinct binding patterns. Protein-DNA interaction was also investigated in vivo, using the Yeast One-Hybrid System. The plasmid constructs were GSTTRI, a DNA fragment composed of three copies of the CCAAT motif of the S. mansoni glutathione S-transferase gene promoter and four oligonucleotides spanning different regions of the S. mansoni p14 gene promoter. None of the yeast clones transformed with the above plasmids was able to grow in selective medium or to activate the transcription of the HIS3 reporter gene, suggesting that SMYB1 could not interact with these promoters in vivo.

Original languageEnglish (US)
Pages (from-to)47-57
Number of pages11
JournalMolecular and Biochemical Parasitology
Volume125
Issue number1-2
DOIs
StatePublished - Jan 1 2002

Keywords

  • CSD
  • EMSA
  • SMYB1
  • Yeast one hybrid system assay

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Y-box binding protein from Schistosoma mansoni: Interaction with DNA and RNA'. Together they form a unique fingerprint.

  • Cite this

    Valadão, A. F., Fantappie, M. R., LoVerde, P. T., Pena, S. D. J., Rumjanek, F. D., & Franco, G. R. (2002). Y-box binding protein from Schistosoma mansoni: Interaction with DNA and RNA. Molecular and Biochemical Parasitology, 125(1-2), 47-57. https://doi.org/10.1016/S0166-6851(02)00210-4