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Bovine CD14 gene characterization and relationship between polymorphisms and surface expression on monocytes and polymorphonuclear neutrophils.

Ibeagha-Awemu EM, Lee JW, Ibeagha AE, Zhao X - BMC Genet. (2008)

Bottom Line: The study has provided information on sequence variations within the CD14 gene and proteins of cattle.Further observations involving a larger sample size are required to validate our findings.The computational analysis on the promoter and comparative analysis with other species has revealed regions of regulatory element motifs that may indicate important regulatory effects on the gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Animal Science, McGill University, Ste-Anne-de-Bellevue, Quebec H9X 3V9, Canada. eveline.ibeagha-awemu@mcgill.ca

ABSTRACT

Background: CD14 is an important player in host innate immunity in that it confers lipopolysaccharide sensitivity to cell types like neutrophils, monocytes and macrophages. The study was aimed at characterizing the CD14 gene of cattle for sequence variations and to determine the effect of variations on the expression of the protein on the surfaces of monocytes and neutrophils in healthy dairy cows.

Results: Five SNPs were identified: two within the coding regions (g.A1908G and g.A2318G, numbering is according to GenBank No. EU148609), one in the 5' (g.C1291T) and two in the 3' (g.A2601G and g.G2621T) untranslated regions. SNP 1908 changes amino acid 175 of the protein (p.Asn175Asp, numbering is according to GenBank No. ABV68569), while SNP 2318 involves a synonymous codon change. Coding region SNPs characterized three gene alleles A (GenBank No. EU148609), A1 (GenBank No. EU148610) and B (GenBank No. EU148611) and two deduced protein variants A (ABV68569 and ABV68570) and B (ABV68571). Protein variant A is more common in the breeds analyzed. All SNPs gave rise to 3 haplotypes for the breeds. SNP genotype 1908AG was significantly (P<0.01) associated with a higher percentage of neutrophils expressing more CD14 molecules on their surfaces. The promoter region contains several transcription factor binding sites, including multiple AP-1 and SP1 sites and there is a high conservation of amino acid residues between the proteins of closely related species.

Conclusion: The study has provided information on sequence variations within the CD14 gene and proteins of cattle. The SNP responsible for an amino acid exchange may play an important role in the expression of CD14 on the surfaces of neutrophils. Further observations involving a larger sample size are required to validate our findings. Our SNP and association analyses have provided baseline information that may be used at defining the role of CD14 in mediating bacterial infections. The computational analysis on the promoter and comparative analysis with other species has revealed regions of regulatory element motifs that may indicate important regulatory effects on the gene.

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Putative transcription factor binding sites (with 100% match) on the bovine CD14 promoter (1213 bps). Recognition sequences are shaded. The rectangle is used where recognition sequences overlap. The arrow points to the first nucleotide of exon 1 at position 1214. Numbering is according to GenBank No. EU148609.
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Figure 2: Putative transcription factor binding sites (with 100% match) on the bovine CD14 promoter (1213 bps). Recognition sequences are shaded. The rectangle is used where recognition sequences overlap. The arrow points to the first nucleotide of exon 1 at position 1214. Numbering is according to GenBank No. EU148609.

Mentions: The CD14 sequence analyzed in this study is made up of 2630 bps with a 1213 bp promoter (Figure 2), two exons and one intron (GenBank No. EU148609). With the use of bioinformatics tools and published information in the literature, we were able to identify putative transcription factor binding sites (TFBSs) (with 100% match against searched data bases) on the bovine CD14 promoter. The putative TFBSs described here are, in particular, those already demonstrated to control the expression of the gene in human and rat. The putative motifs are shown in Figure 2 (boxed) and include amongst others 9 PU.1, 7AP-1, 5 SP1, 4 C/EBP, 3 c-Myb and 2 AP-2 sites.


Bovine CD14 gene characterization and relationship between polymorphisms and surface expression on monocytes and polymorphonuclear neutrophils.

Ibeagha-Awemu EM, Lee JW, Ibeagha AE, Zhao X - BMC Genet. (2008)

Putative transcription factor binding sites (with 100% match) on the bovine CD14 promoter (1213 bps). Recognition sequences are shaded. The rectangle is used where recognition sequences overlap. The arrow points to the first nucleotide of exon 1 at position 1214. Numbering is according to GenBank No. EU148609.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2536669&req=5

Figure 2: Putative transcription factor binding sites (with 100% match) on the bovine CD14 promoter (1213 bps). Recognition sequences are shaded. The rectangle is used where recognition sequences overlap. The arrow points to the first nucleotide of exon 1 at position 1214. Numbering is according to GenBank No. EU148609.
Mentions: The CD14 sequence analyzed in this study is made up of 2630 bps with a 1213 bp promoter (Figure 2), two exons and one intron (GenBank No. EU148609). With the use of bioinformatics tools and published information in the literature, we were able to identify putative transcription factor binding sites (TFBSs) (with 100% match against searched data bases) on the bovine CD14 promoter. The putative TFBSs described here are, in particular, those already demonstrated to control the expression of the gene in human and rat. The putative motifs are shown in Figure 2 (boxed) and include amongst others 9 PU.1, 7AP-1, 5 SP1, 4 C/EBP, 3 c-Myb and 2 AP-2 sites.

Bottom Line: The study has provided information on sequence variations within the CD14 gene and proteins of cattle.Further observations involving a larger sample size are required to validate our findings.The computational analysis on the promoter and comparative analysis with other species has revealed regions of regulatory element motifs that may indicate important regulatory effects on the gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Animal Science, McGill University, Ste-Anne-de-Bellevue, Quebec H9X 3V9, Canada. eveline.ibeagha-awemu@mcgill.ca

ABSTRACT

Background: CD14 is an important player in host innate immunity in that it confers lipopolysaccharide sensitivity to cell types like neutrophils, monocytes and macrophages. The study was aimed at characterizing the CD14 gene of cattle for sequence variations and to determine the effect of variations on the expression of the protein on the surfaces of monocytes and neutrophils in healthy dairy cows.

Results: Five SNPs were identified: two within the coding regions (g.A1908G and g.A2318G, numbering is according to GenBank No. EU148609), one in the 5' (g.C1291T) and two in the 3' (g.A2601G and g.G2621T) untranslated regions. SNP 1908 changes amino acid 175 of the protein (p.Asn175Asp, numbering is according to GenBank No. ABV68569), while SNP 2318 involves a synonymous codon change. Coding region SNPs characterized three gene alleles A (GenBank No. EU148609), A1 (GenBank No. EU148610) and B (GenBank No. EU148611) and two deduced protein variants A (ABV68569 and ABV68570) and B (ABV68571). Protein variant A is more common in the breeds analyzed. All SNPs gave rise to 3 haplotypes for the breeds. SNP genotype 1908AG was significantly (P<0.01) associated with a higher percentage of neutrophils expressing more CD14 molecules on their surfaces. The promoter region contains several transcription factor binding sites, including multiple AP-1 and SP1 sites and there is a high conservation of amino acid residues between the proteins of closely related species.

Conclusion: The study has provided information on sequence variations within the CD14 gene and proteins of cattle. The SNP responsible for an amino acid exchange may play an important role in the expression of CD14 on the surfaces of neutrophils. Further observations involving a larger sample size are required to validate our findings. Our SNP and association analyses have provided baseline information that may be used at defining the role of CD14 in mediating bacterial infections. The computational analysis on the promoter and comparative analysis with other species has revealed regions of regulatory element motifs that may indicate important regulatory effects on the gene.

Show MeSH
Related in: MedlinePlus