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An original SERPINA3 gene cluster: elucidation of genomic organization and gene expression in the Bos taurus 21q24 region.

Pelissier P, Delourme D, Germot A, Blanchet X, Becila S, Maftah A, Leveziel H, Ouali A, Bremaud L - BMC Genomics (2008)

Bottom Line: Finally, in the context of phylogenetic analyses, we improved our knowledge on mammalian SERPINAs evolution.Our experimental results update data of the bovine genome sequencing, substantially increase the bovSERPINA3 sub-family and enrich the phylogenetic tree of serpins.We provide new opportunities for future investigations to approach the biological functions of this unusual subset of serine proteinase inhibitors.

View Article: PubMed Central - HTML - PubMed

Affiliation: INRA, UMR 1061 Unité de Génétique Moléculaire Animale, Université de Limoges, IFR 145, Faculté des Sciences et Techniques, 87060 Limoges, France. patrick.pelissier@unilim.fr

ABSTRACT

Background: The superfamily of serine proteinase inhibitors (serpins) is involved in numerous fundamental biological processes as inflammation, blood coagulation and apoptosis. Our interest is focused on the SERPINA3 sub-family. The major human plasma protease inhibitor, alpha1-antichymotrypsin, encoded by the SERPINA3 gene, is homologous to genes organized in clusters in several mammalian species. However, although there is a similar genic organization with a high degree of sequence conservation, the reactive-centre-loop domains, which are responsible for the protease specificity, show significant divergences.

Results: We provide additional information by analyzing the situation of SERPINA3 in the bovine genome. A cluster of eight genes and one pseudogene sharing a high degree of identity and the same structural organization was characterized. Bovine SERPINA3 genes were localized by radiation hybrid mapping on 21q24 and only spanned over 235 Kilobases. For all these genes, we propose a new nomenclature from SERPINA3-1 to SERPINA3-8. They share approximately 70% of identity with the human SERPINA3 homologue. In the cluster, we described an original sub-group of six members with an unexpected high degree of conservation for the reactive-centre-loop domain, suggesting a similar peptidase inhibitory pattern. Preliminary expression analyses of these bovSERPINA3s showed different tissue-specific patterns and diverse states of glycosylation and phosphorylation. Finally, in the context of phylogenetic analyses, we improved our knowledge on mammalian SERPINAs evolution.

Conclusion: Our experimental results update data of the bovine genome sequencing, substantially increase the bovSERPINA3 sub-family and enrich the phylogenetic tree of serpins. We provide new opportunities for future investigations to approach the biological functions of this unusual subset of serine proteinase inhibitors.

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Southern blot analyses of bovine SERPINA3 organization. (A) Southern blot analysis of bovine genomic DNA digested with three different restriction enzymes NcoI, NciI and SacI. (B) Southern blot analysis of DNA from two overlapping BAC clones bI0123B08 and bI0511D06 digested with the same restriction endonucleases.
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Figure 1: Southern blot analyses of bovine SERPINA3 organization. (A) Southern blot analysis of bovine genomic DNA digested with three different restriction enzymes NcoI, NciI and SacI. (B) Southern blot analysis of DNA from two overlapping BAC clones bI0123B08 and bI0511D06 digested with the same restriction endonucleases.

Mentions: Genomic DNA extracted from bovine blood cells was digested to completion with NciI, NcoI and SacI endonucleases. After transfer on a Hybond-N+ membrane, DNA was hybridized with a radiolabelled probe corresponding to a part of exon 2 of mEndopin 1A gene [GenBank: AY911536]. After washing under high stringency conditions, several fragments of variable intensity were detected for each endonuclease digest as showed in Figure 1A. A minimum of four hybridizing bands was observed for each DNA digestion. In particular, five bands with variable intensity were obtained using SacI. Several fragments were also observed with an additional experiment performed on isolated BAC DNA (see below) treated in the same conditions (Figure 1B). These Southern blot patterns are consistent with the existence of more than the two genes mEndopin1A and mEndopin1B previously described [35]. It suggests a complex organization in the bovine genome with more than a round of duplication during evolution from one ancestral SERPINA3 gene.


An original SERPINA3 gene cluster: elucidation of genomic organization and gene expression in the Bos taurus 21q24 region.

Pelissier P, Delourme D, Germot A, Blanchet X, Becila S, Maftah A, Leveziel H, Ouali A, Bremaud L - BMC Genomics (2008)

Southern blot analyses of bovine SERPINA3 organization. (A) Southern blot analysis of bovine genomic DNA digested with three different restriction enzymes NcoI, NciI and SacI. (B) Southern blot analysis of DNA from two overlapping BAC clones bI0123B08 and bI0511D06 digested with the same restriction endonucleases.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2373789&req=5

Figure 1: Southern blot analyses of bovine SERPINA3 organization. (A) Southern blot analysis of bovine genomic DNA digested with three different restriction enzymes NcoI, NciI and SacI. (B) Southern blot analysis of DNA from two overlapping BAC clones bI0123B08 and bI0511D06 digested with the same restriction endonucleases.
Mentions: Genomic DNA extracted from bovine blood cells was digested to completion with NciI, NcoI and SacI endonucleases. After transfer on a Hybond-N+ membrane, DNA was hybridized with a radiolabelled probe corresponding to a part of exon 2 of mEndopin 1A gene [GenBank: AY911536]. After washing under high stringency conditions, several fragments of variable intensity were detected for each endonuclease digest as showed in Figure 1A. A minimum of four hybridizing bands was observed for each DNA digestion. In particular, five bands with variable intensity were obtained using SacI. Several fragments were also observed with an additional experiment performed on isolated BAC DNA (see below) treated in the same conditions (Figure 1B). These Southern blot patterns are consistent with the existence of more than the two genes mEndopin1A and mEndopin1B previously described [35]. It suggests a complex organization in the bovine genome with more than a round of duplication during evolution from one ancestral SERPINA3 gene.

Bottom Line: Finally, in the context of phylogenetic analyses, we improved our knowledge on mammalian SERPINAs evolution.Our experimental results update data of the bovine genome sequencing, substantially increase the bovSERPINA3 sub-family and enrich the phylogenetic tree of serpins.We provide new opportunities for future investigations to approach the biological functions of this unusual subset of serine proteinase inhibitors.

View Article: PubMed Central - HTML - PubMed

Affiliation: INRA, UMR 1061 Unité de Génétique Moléculaire Animale, Université de Limoges, IFR 145, Faculté des Sciences et Techniques, 87060 Limoges, France. patrick.pelissier@unilim.fr

ABSTRACT

Background: The superfamily of serine proteinase inhibitors (serpins) is involved in numerous fundamental biological processes as inflammation, blood coagulation and apoptosis. Our interest is focused on the SERPINA3 sub-family. The major human plasma protease inhibitor, alpha1-antichymotrypsin, encoded by the SERPINA3 gene, is homologous to genes organized in clusters in several mammalian species. However, although there is a similar genic organization with a high degree of sequence conservation, the reactive-centre-loop domains, which are responsible for the protease specificity, show significant divergences.

Results: We provide additional information by analyzing the situation of SERPINA3 in the bovine genome. A cluster of eight genes and one pseudogene sharing a high degree of identity and the same structural organization was characterized. Bovine SERPINA3 genes were localized by radiation hybrid mapping on 21q24 and only spanned over 235 Kilobases. For all these genes, we propose a new nomenclature from SERPINA3-1 to SERPINA3-8. They share approximately 70% of identity with the human SERPINA3 homologue. In the cluster, we described an original sub-group of six members with an unexpected high degree of conservation for the reactive-centre-loop domain, suggesting a similar peptidase inhibitory pattern. Preliminary expression analyses of these bovSERPINA3s showed different tissue-specific patterns and diverse states of glycosylation and phosphorylation. Finally, in the context of phylogenetic analyses, we improved our knowledge on mammalian SERPINAs evolution.

Conclusion: Our experimental results update data of the bovine genome sequencing, substantially increase the bovSERPINA3 sub-family and enrich the phylogenetic tree of serpins. We provide new opportunities for future investigations to approach the biological functions of this unusual subset of serine proteinase inhibitors.

Show MeSH