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Originally published In Press as doi:10.1074/jbc.M909547199 on May 5, 2000

J. Biol. Chem., Vol. 275, Issue 31, 23861-23868, August 4, 2000
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Promoter Sequences of the Putative Anopheles gambiae Apyrase Confer Salivary Gland Expression in Drosophila melanogaster*

Fabrizio LombardoDagger ||§, Manlio Di Cristina, Lefteris Spanos||, Christos Louis||**, Mario ColuzziDagger , and Bruno ArcàDagger Dagger Dagger §§

From the Dagger  Istituto di Parassitologia, Istituto Pasteur-Fondazione Cenci Bolognetti, Università di Roma "La Sapienza," 00185 Roma, Italy,  Department of Biology, Imperial College of Science, Technology, and Medicine, London SW7 2AZ, United Kingdom, || Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, 71110 Heraklion, Crete, Greece, ** Department of Biology, University of Crete, 71110 Heraklion, Crete, Greece, and Dagger Dagger  Dipartimento di Genetica, Biologia Generale e Molecolare, Università di Napoli Federico II, 80134 Napoli, Italy

The saliva of blood-feeding arthropods contains an apyrase that facilitates hematophagy by inhibiting the ADP-induced aggregation of the host platelets. We report here the isolation of a salivary gland-specific cDNA encoding a secreted protein that likely represents the Anopheles gambiae apyrase. We describe also two additional members of the apyrase/5'-nucleotidase family. The cDNA corresponding to the AgApyL1 gene encodes a secreted protein that is closely related in sequence to the apyrase of the yellow fever mosquito, Aedes aegypti, and whose expression appears enriched in, but not restricted to, female salivary glands. The AgApyL2 gene was found searching an A. gambiae data base, and its expression is restricted to larval stages. We isolated the gene encoding the presumed A. gambiae apyrase (AgApy) and we tested its putative promoter for the tissue-specific expression of the LacZ gene from Escherichia coli in transgenic Drosophila melanogaster. All the transgenic lines analyzed showed a weak but unambiguous staining of the adult glands, indicating that some of the salivary gland-specific transcriptional regulatory elements are conserved between the malaria mosquito and the fruit fly. The availability of salivary gland-specific promoters may be useful both for studies on vector-parasite interactions and, potentially, for the targeted tissue-specific expression of anti-parasite genes in the mosquito.

* This work was supported by European Union Grant ERBFMRXCT960017 (to M. C and C. L.) and by a grant from the United Nations Developmental Program/World Bank/World Health Organization Special Program for Research and Training in Tropical Diseases (TDR) (to M. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AJ237704, AJ237705, and AJ237706.

§ Supported by a training fellowship of the University of Rome "La Sapienza."

§§ Supported by a postdoctoral fellowship of the Istituto Pasteur-Fondazione Cenci Bolognetti and by European Union Return Grant BIO4-CT98-5020. To whom correspondence should be addressed: Istituto di Parassitologia, Università di Roma "La Sapienza," P. le Aldo Moro 5, Box 6, Roma 62, 00185 Roma, Italy. Tel.: 39-06-4991-4900; Fax: 39-06-4991-4644; E-mail: b.Arca@Caspur.it.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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