Identification and functional characterization of a member of the PUR-α family from Schistosoma mansoni

Marcelo R. Fantappie, Ahmed Osman, Edward G. Niles, Philip T. LoVerde

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Schistosoma mansoni p14 gene encodes an eggshell precursor that is expressed only in vitelline cells of mature female worms in response to a male stimulus. The upstream region of the p14 gene contains several potential cis-acting regulatory sequences. We used the upstream region of the p14 gene as bait in a yeast-one-hybrid screen of a S. mansoni cDNA library to identify interacting proteins. We report the identification and characterization of a cDNA (S. mansoni PUR-alpha (SmPUR-α)) encoding a protein homologous to single-stranded DNA transcription activator PUR-α, that binds to the p14 upstream region and activates transcription of the HIS3 reporter gene in yeast. SmPUR-α has a predicted molecular mass of 30 kDa and shares an overall homology of 63% with mammalian PUR-α. The DNA binding domain of SmPUR-α is highly conserved. We show by gel shift assays that GST-SmPUR-α binds to oligonucleotides comprising the p14 upstream region. SmPUR-α binds preferentially to single-stranded DNA and also binds RNA. Unlike the mammalian homologue, SmPUR-α exhibits little specificity for the PUR element GGn, but shows strong preference for a sequence containing alternating pyrimidines. Our data support that SmPUR-α is a single-copy gene and through reverse transcriptase-polymerase chain reaction and in situ hybridization, we show that SmPUR-α is constitutively transcribed in many cell types and thus likely plays a role as a general transcription activator in schistosomes. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)373-390
Number of pages18
JournalMolecular and Biochemical Parasitology
Volume110
Issue number2
DOIs
StatePublished - 2000

Keywords

  • PUR-alpha
  • Schistosoma mansoni
  • Single-stranded DNA binding protein
  • Transcription regulation

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Identification and functional characterization of a member of the PUR-α family from Schistosoma mansoni'. Together they form a unique fingerprint.

Cite this