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Infection and Immunity, May 2003, p. 2548-2554, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2548-2554.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Isolation of cDNAs Encoding Secreted and Transmembrane Proteins from Schistosoma mansoni by a Signal Sequence Trap Method

Danielle Smyth,1 Donald P. McManus,1 Michael J. Smout,1,2 Thewarach Laha,1 Wenbao Zhang,1 and Alex Loukas1,2*

Molecular Parasitology Laboratory, Australian Centre for International and Tropical Health and Nutrition, The Queensland Institute of Medical Research and The University of Queensland, Brisbane, Queensland 4029, Australia,1 Department of Microbiology & Tropical Medicine, The George Washington University Medical Center, Washington, D.C.2

Received 1 October 2002/ Returned for modification 25 November 2002/ Accepted 24 January 2003

Surface and secreted proteins of schistosomes orchestrate the basic physiologic requirements of a parasitic existence. These proteins are often exposed to host tissues during penetration, migration, feeding, and immune evasion, and they are obvious targets for control strategies. Signal sequence trap (SST) represents a novel approach that selects for cDNAs encoding secreted and surface proteins with N-terminal signal peptides, so we constructed a randomly primed adult Schistosoma mansoni cDNA library fused to a signalless reporter gene encoding placental alkaline phosphatase. The library was used to transfect COS-7 cells, which were then assayed for the presence of reporter at the cell surface. Eighteen S. mansoni cDNA fragments were isolated and sequenced. Expression profiles of the novel clones were determined for different developmental stages; some transcripts were restricted to single-sex adult worms, while others were ubiquitously distributed. Most clones contained signal peptides or signal anchors as determined by the SignalP algorithm. Open reading frames (ORFs) were categorized as follows: (i) previously identified S. mansoni cDNAs encoding proteins of known function; (ii) cDNAs encoding proteins of known function in other organisms but novel for Schistosoma; (iii) S. mansoni expressed sequence tags (ESTs) of unknown function; and (iv) completely novel ORFs without homologues (including ESTs) from any phylum. Clones of particular interest included tetraspanins similar to human cell surface antigens, a protein kinase, and ORFs transcribed in the antisense orientation to previously characterized S. mansoni cDNAs. This is the first report describing the use of SST as a tool for identifying secreted proteins from any pathogenic organism.

* Corresponding author. Mailing address: Department of Microbiology & Tropical Medicine, Ross Hall, Room 726, The George Washington University Medical Center, 2300 Eye St., NW, Washington, D.C. 20037. Phone: (202) 994-2150. Fax: (202) 994-2913. E-mail: mtmacl{at}gwumc.edu.

Editor: W. A. Petri, Jr.

Infection and Immunity, May 2003, p. 2548-2554, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2548-2554.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • McManus, D. P., Loukas, A. (2008). Current Status of Vaccines for Schistosomiasis. Clin. Microbiol. Rev. 21: 225-242 [Abstract] [Full Text]  
  • Knudsen, G. M., Medzihradszky, K. F., Lim, K.-C., Hansell, E., McKerrow, J. H. (2005). Proteomic Analysis of Schistosoma mansoni Cercarial Secretions. Mol. Cell. Proteomics 4: 1862-1875 [Abstract] [Full Text]  


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