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Evolution of an expanded mannose receptor gene family.

Staines K, Hunt LG, Young JR, Butter C - PLoS ONE (2014)

Bottom Line: Inspection of the genomic environment of the chicken gene revealed an array of five paralogous genes, MRC1L-A to MRC1L-E, located between conserved flanking genes found either side of the single MRC1 gene in mammals.The lizard genome had only three genes, one orthologue of MRC1L-A and two orthologues of the MRC1L-B antigen gene resulting from a recent duplication.Cytoplasmic regions of the other three genes were very divergent, possibly indicating the evolution of a new functional repertoire for this family of molecules, which might include novel interactions with pathogens.

View Article: PubMed Central - PubMed

Affiliation: The Pirbright Institute, Compton, United Kingdom.

ABSTRACT
Sequences of peptides from a protein specifically immunoprecipitated by an antibody, KUL01, that recognises chicken macrophages, identified a homologue of the mammalian mannose receptor, MRC1, which we called MRC1L-B. Inspection of the genomic environment of the chicken gene revealed an array of five paralogous genes, MRC1L-A to MRC1L-E, located between conserved flanking genes found either side of the single MRC1 gene in mammals. Transcripts of all five genes were detected in RNA from a macrophage cell line and other RNAs, whose sequences allowed the precise definition of spliced exons, confirming or correcting existing bioinformatic annotation. The confirmed gene structures were used to locate orthologues of all five genes in the genomes of two other avian species and of the painted turtle, all with intact coding sequences. The lizard genome had only three genes, one orthologue of MRC1L-A and two orthologues of the MRC1L-B antigen gene resulting from a recent duplication. The Xenopus genome, like that of most mammals, had only a single MRC1-like gene at the corresponding locus. MRC1L-A and MRC1L-B genes had similar cytoplasmic regions that may be indicative of similar subcellular migration and functions. Cytoplasmic regions of the other three genes were very divergent, possibly indicating the evolution of a new functional repertoire for this family of molecules, which might include novel interactions with pathogens.

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KUL01 specifically precipitates a molecule with apparent molecular weight 180 kDa.Track M contains molecular weight standards. The other tracks contain materials absorbed from a precleared lysate of the HD11 macrophage cell line, by agarose beads to which were attached either KUL01, an isotype matched control antibody, or no antibody, and eluted at low pH. The open arrowhead points to the band(s) specifically absorbed by the KUL01 antibody, which were analysed by mass spectroscopy.
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pone-0110330-g001: KUL01 specifically precipitates a molecule with apparent molecular weight 180 kDa.Track M contains molecular weight standards. The other tracks contain materials absorbed from a precleared lysate of the HD11 macrophage cell line, by agarose beads to which were attached either KUL01, an isotype matched control antibody, or no antibody, and eluted at low pH. The open arrowhead points to the band(s) specifically absorbed by the KUL01 antibody, which were analysed by mass spectroscopy.

Mentions: KUL01 antibody bound to agarose beads was used to adsorb proteins from a lysate of the transformed chicken macrophage cell line HD11, which were analysed by SDS PAGE after elution at low pH. Specific bands, obtained from beads coated with KUL01 but not from those coated with control antibody, were excised, digested with trypsin and analysed my mass spectroscopy. The major specifically recognised molecule was a (doublet) band with an apparent molecular weight of 180 kDa (figure 1). By Mascot search of the NCBI non-redundant chicken proteins in the IPI database, a sufficient number of peptides from the tryptic digest of this band were identified as being derivable from the sequence IPI00814304 to unequivocally identify it as the source of antigen specifically adsorbed by KUL01 (figure S1 and table S1). It was annotated as being a chicken homologue of MRC1.


Evolution of an expanded mannose receptor gene family.

Staines K, Hunt LG, Young JR, Butter C - PLoS ONE (2014)

KUL01 specifically precipitates a molecule with apparent molecular weight 180 kDa.Track M contains molecular weight standards. The other tracks contain materials absorbed from a precleared lysate of the HD11 macrophage cell line, by agarose beads to which were attached either KUL01, an isotype matched control antibody, or no antibody, and eluted at low pH. The open arrowhead points to the band(s) specifically absorbed by the KUL01 antibody, which were analysed by mass spectroscopy.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4229073&req=5

pone-0110330-g001: KUL01 specifically precipitates a molecule with apparent molecular weight 180 kDa.Track M contains molecular weight standards. The other tracks contain materials absorbed from a precleared lysate of the HD11 macrophage cell line, by agarose beads to which were attached either KUL01, an isotype matched control antibody, or no antibody, and eluted at low pH. The open arrowhead points to the band(s) specifically absorbed by the KUL01 antibody, which were analysed by mass spectroscopy.
Mentions: KUL01 antibody bound to agarose beads was used to adsorb proteins from a lysate of the transformed chicken macrophage cell line HD11, which were analysed by SDS PAGE after elution at low pH. Specific bands, obtained from beads coated with KUL01 but not from those coated with control antibody, were excised, digested with trypsin and analysed my mass spectroscopy. The major specifically recognised molecule was a (doublet) band with an apparent molecular weight of 180 kDa (figure 1). By Mascot search of the NCBI non-redundant chicken proteins in the IPI database, a sufficient number of peptides from the tryptic digest of this band were identified as being derivable from the sequence IPI00814304 to unequivocally identify it as the source of antigen specifically adsorbed by KUL01 (figure S1 and table S1). It was annotated as being a chicken homologue of MRC1.

Bottom Line: Inspection of the genomic environment of the chicken gene revealed an array of five paralogous genes, MRC1L-A to MRC1L-E, located between conserved flanking genes found either side of the single MRC1 gene in mammals.The lizard genome had only three genes, one orthologue of MRC1L-A and two orthologues of the MRC1L-B antigen gene resulting from a recent duplication.Cytoplasmic regions of the other three genes were very divergent, possibly indicating the evolution of a new functional repertoire for this family of molecules, which might include novel interactions with pathogens.

View Article: PubMed Central - PubMed

Affiliation: The Pirbright Institute, Compton, United Kingdom.

ABSTRACT
Sequences of peptides from a protein specifically immunoprecipitated by an antibody, KUL01, that recognises chicken macrophages, identified a homologue of the mammalian mannose receptor, MRC1, which we called MRC1L-B. Inspection of the genomic environment of the chicken gene revealed an array of five paralogous genes, MRC1L-A to MRC1L-E, located between conserved flanking genes found either side of the single MRC1 gene in mammals. Transcripts of all five genes were detected in RNA from a macrophage cell line and other RNAs, whose sequences allowed the precise definition of spliced exons, confirming or correcting existing bioinformatic annotation. The confirmed gene structures were used to locate orthologues of all five genes in the genomes of two other avian species and of the painted turtle, all with intact coding sequences. The lizard genome had only three genes, one orthologue of MRC1L-A and two orthologues of the MRC1L-B antigen gene resulting from a recent duplication. The Xenopus genome, like that of most mammals, had only a single MRC1-like gene at the corresponding locus. MRC1L-A and MRC1L-B genes had similar cytoplasmic regions that may be indicative of similar subcellular migration and functions. Cytoplasmic regions of the other three genes were very divergent, possibly indicating the evolution of a new functional repertoire for this family of molecules, which might include novel interactions with pathogens.

Show MeSH