Isolation and characterization of a new chemokine receptor gene, the putative chicken CXCR1.
This study delineates the isolation and characterization of a novel chemokine receptor gene, the putative chicken CXC receptor 1 (cCXCR1). Using a human CXCR1 probe, we isolated several positive clones from a chicken genomic library. One of the clones contained a fragment of approximately 5000bp that hybridized strongly with the hCXCR1 probe. This fragment was sequenced and subjected to a variety of computer analyses. The open reading frame for this gene predicts a seven transmembrane domain protein with all the characteristics of a chemokine receptor and with 67% sequence homology to hCXCR1, 65% to hCXCR2 and also with considerable sequence homology to other human chemokine receptors such as hCXCR4 (50%), hCCR2 (49%) and hCCR1 (49%). However, the homology to a previously isolated potential G-protein-coupled receptor for chickens (AvCRL1) is only 47%. Using 5' RACE, two transcription initiation sites were identified suggesting the potential for the expression of two protein isoforms (I and II) in vivo. The promoter for the putative cCXCR1 contains a variety of consensus transcription factor binding elements that can potentially be involved in the expression of this chicken receptor upon stimulation by stress-inducing agents. RT-PCR analysis was used to determine the pattern of expression of the larger isoform (I) of this receptor in a variety of tissues. This form of the receptor is expressed primarily in the organs of the gastrointestinal tract, tissues that are frequently exposed to stress-inducing agents, but not in the central nervous system, tissues that are protected from insult by the blood barrier. Using the same RT-PCR approach we show that stress-inducing agents, such as 'first-hand' and 'second-hand' cigarette smoke components, tumor promoters and thrombin, differentially stimulate the expression of the isoform I in primary fibroblasts. Thrombin is an enzyme that plays many important roles in thrombosis, angiogenesis and wound healing and exposure to both cigarette smokes and/or to tumor promoters can lead to tumorigenesis. Therefore, upregulation of chemokines and their receptors by stress-inducing agents can confer highly regulated modulation of cellular responses to traumatic and pathological situations.
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Interdepartmental Graduate Program in Biochemistry, University of California, Riverside, CA 92521, USA.
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Martins-Green
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation / Q000187:drug effects; D005796:Genes / Q000235:genetics*; D008969:Molecular Sequence Data; D010703:Phorbol Esters / Q000494:pharmacology; D020033:Protein Isoforms / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism; D019707:Receptors, Chemokine / Q000235:genetics*; D023062:Receptors, Interleukin-8A / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013917:Thrombin / Q000494:pharmacology; D014018:Tissue Distribution; D014158:Transcription, Genetic
D010703:Phorbol Esters; D020033:Protein Isoforms; D019707:Receptors, Chemokine; D023062:Receptors, Interleukin-8A; D012313:RNA; D004247:DNA; D013917:Thrombin