TY - JOUR
T1 - Isolation of a functional ecdysteroid receptor homologue from the ixodid tick Amblyomma americanum (L.)
AU - Guo, Xiaoping
AU - Harmon, Margaret A.
AU - Laudet, Vincent
AU - Mangelsdorf, David J.
AU - Palmer, Melanie J.
N1 - Funding Information:
The authors would like to thank the Oklahoma State University Recombinant DNA/Protein Resource Facility for the synthesis and purification of synthetic oligonucleotides and for automated DNA sequencing. We would also like to thank Dr Alexander Raikhel for suggestions about designing degenerate EcR primers and the laboratory of Dr David Hogness for the DmEcR-17 cDNA. We would like to acknowledge the laboratory of Dr Jean Delachambre for sharing T. molitor EcR results prior to publication, Dr Roberta Kelleher for salivary gland and ovary dissections, and Drs John Sauer and Stephen Wikel for providing comments on the manuscript. This work was supported by Oklahoma Advancement of Science and Technology (OCAST) (HR-016), National Research Initiative Competitive Grants Program (95-37302-2360), and Oklahoma Agricultural Experiment Station Hatch (OKLO2139) grants to MJP, Howard Hughes Medical Institute (HHMI) and a Robert A. Welch Foundation Grant to DJM, and support from CNRS and the Institut Pasteur de Lille to VL.
PY - 1998/11
Y1 - 1998/11
N2 - Ecdysteroids are assumed to be the major steroid hormones in arthropods. However, with the exception of insects and crustaceans, very little is known about ecdysteroid action in other arthropods. To determine if ecdysteriods play a functional role in the ixodid tick, Amblyomma americanum (L.), we isolated cDNAs encoding three presumed ecdysteroid receptor isoforms (AamEcRA1, AamEcRA2, and AamEcRA3) that have common DNA and ligand binding domains linked to distinct amino termini. The DNA and ligand binding domains share an average of 86 and 64% identity, respectively with DNA and ligand binding domains from insect EcR proteins. The amino termini are highly divergent and the AamEcRs lack the 'F' domain found in the insect EcRs. Analysis of AamEcR cDNAs show that processing of the AamEcR gene is complex, producing multiple transcripts with unique 5' and 3' termini as well as splicing variants with incomplete open reading frames. AamEcR mRNA profiles in whole animals and isolated tissues are consistent with complex regulation of AamEcR expression. We also examined the ability of AamEcRA1, when paired with an AamRXR, to activate transcription of an ecdysone response element containing reporter, and demonstrate that the AamEcR gene encodes a functional ecdysteroid receptor.
AB - Ecdysteroids are assumed to be the major steroid hormones in arthropods. However, with the exception of insects and crustaceans, very little is known about ecdysteroid action in other arthropods. To determine if ecdysteriods play a functional role in the ixodid tick, Amblyomma americanum (L.), we isolated cDNAs encoding three presumed ecdysteroid receptor isoforms (AamEcRA1, AamEcRA2, and AamEcRA3) that have common DNA and ligand binding domains linked to distinct amino termini. The DNA and ligand binding domains share an average of 86 and 64% identity, respectively with DNA and ligand binding domains from insect EcR proteins. The amino termini are highly divergent and the AamEcRs lack the 'F' domain found in the insect EcRs. Analysis of AamEcR cDNAs show that processing of the AamEcR gene is complex, producing multiple transcripts with unique 5' and 3' termini as well as splicing variants with incomplete open reading frames. AamEcR mRNA profiles in whole animals and isolated tissues are consistent with complex regulation of AamEcR expression. We also examined the ability of AamEcRA1, when paired with an AamRXR, to activate transcription of an ecdysone response element containing reporter, and demonstrate that the AamEcR gene encodes a functional ecdysteroid receptor.
KW - Arnblyomma americanum
KW - Ecdysone
KW - Ecdysteroid receptor
KW - Tick
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U2 - 10.1016/S0965-1748(97)00075-1
DO - 10.1016/S0965-1748(97)00075-1
M3 - Article
C2 - 9501418
AN - SCOPUS:0344382060
SN - 0965-1748
VL - 27
SP - 945
EP - 962
JO - Insect Biochemistry and Molecular Biology
JF - Insect Biochemistry and Molecular Biology
IS - 11
ER -