Molecular cloning, functional expression, and gene silencing of two Drosophila receptors for the Drosophila neuropeptide pyrokinin-2
The database of the Drosophila Genome Project contains the sequences of two genes, CG8784 and CG8795, predicted to code for two structurally related G protein-coupled receptors. We have cloned these genes and expressed their coding parts in Chinese hamster ovary cells. We found that both receptors c...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2003-09, Vol.309 (2), p.485-494 |
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| creator | Rosenkilde, Carina Cazzamali, Giuseppe Williamson, Michael Hauser, Frank Søndergaard, Leif DeLotto, Robert Grimmelikhuijzen, Cornelis J.P |
| description | The database of the
Drosophila Genome Project contains the sequences of two genes, CG8784 and CG8795, predicted to code for two structurally related G protein-coupled receptors. We have cloned these genes and expressed their coding parts in Chinese hamster ovary cells. We found that both receptors can be activated by low concentrations of the
Drosophila neuropeptide pyrokinin-2 (CG8784, EC
50 for pyrokinin-2, 1
×
10
−9
M; CG8795, EC
50 for pyrokinin-2, 5
×
10
−10
M). The precise role of
Drosophila pyrokinin-2 (SVPFKPRLamide) in
Drosophila is unknown, but in other insects, pyrokinins have diverse myotropic actions and are also initiating sex pheromone biosynthesis and embryonic diapause. Gene silencing, using the RNA-mediated interference technique, showed that CG8784 gene silencing caused lethality in embryos, whereas CG8795 gene silencing resulted in strongly reduced viability for both embryos and first instar larvae. In addition to the two
Drosophila receptors, we also identified two probable pyrokinin receptors in the genomic database from the malaria mosquito
Anopheles gambiae. The two
Drosophila pyrokinin receptors are, to our knowledge, the first invertebrate pyrokinin receptors to be identified. |
| doi_str_mv | 10.1016/j.bbrc.2003.08.022 |
| format | Article |
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Drosophila Genome Project contains the sequences of two genes, CG8784 and CG8795, predicted to code for two structurally related G protein-coupled receptors. We have cloned these genes and expressed their coding parts in Chinese hamster ovary cells. We found that both receptors can be activated by low concentrations of the
Drosophila neuropeptide pyrokinin-2 (CG8784, EC
50 for pyrokinin-2, 1
×
10
−9
M; CG8795, EC
50 for pyrokinin-2, 5
×
10
−10
M). The precise role of
Drosophila pyrokinin-2 (SVPFKPRLamide) in
Drosophila is unknown, but in other insects, pyrokinins have diverse myotropic actions and are also initiating sex pheromone biosynthesis and embryonic diapause. Gene silencing, using the RNA-mediated interference technique, showed that CG8784 gene silencing caused lethality in embryos, whereas CG8795 gene silencing resulted in strongly reduced viability for both embryos and first instar larvae. In addition to the two
Drosophila receptors, we also identified two probable pyrokinin receptors in the genomic database from the malaria mosquito
Anopheles gambiae. The two
Drosophila pyrokinin receptors are, to our knowledge, the first invertebrate pyrokinin receptors to be identified.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2003.08.022</identifier><identifier>PMID: 12951076</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Anopheles gambiae ; CG8784 gene ; CG8795 gene ; CHO Cells - metabolism ; Cloning, Molecular ; Cricetinae - genetics ; Cricetinae - metabolism ; Drosophila ; Drosophila - genetics ; Drosophila - metabolism ; Gene Expression Regulation ; Gene Silencing - drug effects ; GTP-Binding Proteins - chemistry ; GTP-Binding Proteins - classification ; GTP-Binding Proteins - genetics ; GTP-Binding Proteins - metabolism ; Molecular Sequence Data ; Neuropeptides - genetics ; Neuropeptides - metabolism ; pyrokinins ; Sequence Alignment ; Sequence Analysis, Protein ; Transfection</subject><ispartof>Biochemical and biophysical research communications, 2003-09, Vol.309 (2), p.485-494</ispartof><rights>2003 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c449t-bae31a16c8c10f6e099732fcfa6c5bb66cf888cb5652a6ce088b2508729f88933</citedby><cites>FETCH-LOGICAL-c449t-bae31a16c8c10f6e099732fcfa6c5bb66cf888cb5652a6ce088b2508729f88933</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0006291X03016139$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12951076$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rosenkilde, Carina</creatorcontrib><creatorcontrib>Cazzamali, Giuseppe</creatorcontrib><creatorcontrib>Williamson, Michael</creatorcontrib><creatorcontrib>Hauser, Frank</creatorcontrib><creatorcontrib>Søndergaard, Leif</creatorcontrib><creatorcontrib>DeLotto, Robert</creatorcontrib><creatorcontrib>Grimmelikhuijzen, Cornelis J.P</creatorcontrib><title>Molecular cloning, functional expression, and gene silencing of two Drosophila receptors for the Drosophila neuropeptide pyrokinin-2</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>The database of the
Drosophila Genome Project contains the sequences of two genes, CG8784 and CG8795, predicted to code for two structurally related G protein-coupled receptors. We have cloned these genes and expressed their coding parts in Chinese hamster ovary cells. We found that both receptors can be activated by low concentrations of the
Drosophila neuropeptide pyrokinin-2 (CG8784, EC
50 for pyrokinin-2, 1
×
10
−9
M; CG8795, EC
50 for pyrokinin-2, 5
×
10
−10
M). The precise role of
Drosophila pyrokinin-2 (SVPFKPRLamide) in
Drosophila is unknown, but in other insects, pyrokinins have diverse myotropic actions and are also initiating sex pheromone biosynthesis and embryonic diapause. Gene silencing, using the RNA-mediated interference technique, showed that CG8784 gene silencing caused lethality in embryos, whereas CG8795 gene silencing resulted in strongly reduced viability for both embryos and first instar larvae. In addition to the two
Drosophila receptors, we also identified two probable pyrokinin receptors in the genomic database from the malaria mosquito
Anopheles gambiae. The two
Drosophila pyrokinin receptors are, to our knowledge, the first invertebrate pyrokinin receptors to be identified.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Anopheles gambiae</subject><subject>CG8784 gene</subject><subject>CG8795 gene</subject><subject>CHO Cells - metabolism</subject><subject>Cloning, Molecular</subject><subject>Cricetinae - genetics</subject><subject>Cricetinae - metabolism</subject><subject>Drosophila</subject><subject>Drosophila - genetics</subject><subject>Drosophila - metabolism</subject><subject>Gene Expression Regulation</subject><subject>Gene Silencing - drug effects</subject><subject>GTP-Binding Proteins - chemistry</subject><subject>GTP-Binding Proteins - classification</subject><subject>GTP-Binding Proteins - genetics</subject><subject>GTP-Binding Proteins - metabolism</subject><subject>Molecular Sequence Data</subject><subject>Neuropeptides - genetics</subject><subject>Neuropeptides - metabolism</subject><subject>pyrokinins</subject><subject>Sequence Alignment</subject><subject>Sequence Analysis, Protein</subject><subject>Transfection</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2OFCEUhYnROO3oC7gwrFxNlReqiobEjRl_kzFuNHFHKOoyQ0tDCVWjs_fBpdOd6EpXBM53zw3nEPKUQcuAiRe7dhyzbTlA14JsgfN7ZMNAQcMZ9PfJBgBEwxX7ekYelbIDYKwX6iE5Y1wNDLZiQ359TAHtGkymNqTo4_UFdWu0i0_RBIo_54yl1MsFNXGi1xiRFh8w2orS5OjyI9HXOZU03_hgaEaL85JyoS5lutzg32LENae56n5COt_l9M3XjQ1_TB44Ewo-OZ3n5MvbN58v3zdXn959uHx11di-V0szGuyYYcJKy8AJBKW2HXfWGWGHcRTCOimlHQcx8PqEIOXIB5Bbrqqguu6cPD_6zjl9X7Eseu-LxRBMxLQWve1ETWXo_wsyqXqQHasgP4K2_rJkdHrOfm_ynWagDyXpnT6UpA8laZC6llSHnp3c13GP05-RUysVeHkEsIZx6zHrYn2NHCdf8130lPy__H8D4HWlnQ</recordid><startdate>20030919</startdate><enddate>20030919</enddate><creator>Rosenkilde, Carina</creator><creator>Cazzamali, Giuseppe</creator><creator>Williamson, Michael</creator><creator>Hauser, Frank</creator><creator>Søndergaard, Leif</creator><creator>DeLotto, Robert</creator><creator>Grimmelikhuijzen, Cornelis J.P</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SS</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20030919</creationdate><title>Molecular cloning, functional expression, and gene silencing of two Drosophila receptors for the Drosophila neuropeptide pyrokinin-2</title><author>Rosenkilde, Carina ; Cazzamali, Giuseppe ; Williamson, Michael ; Hauser, Frank ; Søndergaard, Leif ; DeLotto, Robert ; Grimmelikhuijzen, Cornelis J.P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c449t-bae31a16c8c10f6e099732fcfa6c5bb66cf888cb5652a6ce088b2508729f88933</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Anopheles gambiae</topic><topic>CG8784 gene</topic><topic>CG8795 gene</topic><topic>CHO Cells - metabolism</topic><topic>Cloning, Molecular</topic><topic>Cricetinae - genetics</topic><topic>Cricetinae - metabolism</topic><topic>Drosophila</topic><topic>Drosophila - genetics</topic><topic>Drosophila - metabolism</topic><topic>Gene Expression Regulation</topic><topic>Gene Silencing - drug effects</topic><topic>GTP-Binding Proteins - chemistry</topic><topic>GTP-Binding Proteins - classification</topic><topic>GTP-Binding Proteins - genetics</topic><topic>GTP-Binding Proteins - metabolism</topic><topic>Molecular Sequence Data</topic><topic>Neuropeptides - genetics</topic><topic>Neuropeptides - metabolism</topic><topic>pyrokinins</topic><topic>Sequence Alignment</topic><topic>Sequence Analysis, Protein</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rosenkilde, Carina</creatorcontrib><creatorcontrib>Cazzamali, Giuseppe</creatorcontrib><creatorcontrib>Williamson, Michael</creatorcontrib><creatorcontrib>Hauser, Frank</creatorcontrib><creatorcontrib>Søndergaard, Leif</creatorcontrib><creatorcontrib>DeLotto, Robert</creatorcontrib><creatorcontrib>Grimmelikhuijzen, Cornelis J.P</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rosenkilde, Carina</au><au>Cazzamali, Giuseppe</au><au>Williamson, Michael</au><au>Hauser, Frank</au><au>Søndergaard, Leif</au><au>DeLotto, Robert</au><au>Grimmelikhuijzen, Cornelis J.P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning, functional expression, and gene silencing of two Drosophila receptors for the Drosophila neuropeptide pyrokinin-2</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2003-09-19</date><risdate>2003</risdate><volume>309</volume><issue>2</issue><spage>485</spage><epage>494</epage><pages>485-494</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>The database of the
Drosophila Genome Project contains the sequences of two genes, CG8784 and CG8795, predicted to code for two structurally related G protein-coupled receptors. We have cloned these genes and expressed their coding parts in Chinese hamster ovary cells. We found that both receptors can be activated by low concentrations of the
Drosophila neuropeptide pyrokinin-2 (CG8784, EC
50 for pyrokinin-2, 1
×
10
−9
M; CG8795, EC
50 for pyrokinin-2, 5
×
10
−10
M). The precise role of
Drosophila pyrokinin-2 (SVPFKPRLamide) in
Drosophila is unknown, but in other insects, pyrokinins have diverse myotropic actions and are also initiating sex pheromone biosynthesis and embryonic diapause. Gene silencing, using the RNA-mediated interference technique, showed that CG8784 gene silencing caused lethality in embryos, whereas CG8795 gene silencing resulted in strongly reduced viability for both embryos and first instar larvae. In addition to the two
Drosophila receptors, we also identified two probable pyrokinin receptors in the genomic database from the malaria mosquito
Anopheles gambiae. The two
Drosophila pyrokinin receptors are, to our knowledge, the first invertebrate pyrokinin receptors to be identified.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>12951076</pmid><doi>10.1016/j.bbrc.2003.08.022</doi><tpages>10</tpages></addata></record> |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Amino Acid Sequence Animals Anopheles gambiae CG8784 gene CG8795 gene CHO Cells - metabolism Cloning, Molecular Cricetinae - genetics Cricetinae - metabolism Drosophila Drosophila - genetics Drosophila - metabolism Gene Expression Regulation Gene Silencing - drug effects GTP-Binding Proteins - chemistry GTP-Binding Proteins - classification GTP-Binding Proteins - genetics GTP-Binding Proteins - metabolism Molecular Sequence Data Neuropeptides - genetics Neuropeptides - metabolism pyrokinins Sequence Alignment Sequence Analysis, Protein Transfection |
| title | Molecular cloning, functional expression, and gene silencing of two Drosophila receptors for the Drosophila neuropeptide pyrokinin-2 |
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