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Functional characterization of a BCL10 isoform in the rainbow trout Oncorhynchus mykiss.

Mazzone P, Scudiero I, Coccia E, Ferravante A, Paolucci M, D'Andrea EL, Varricchio E, Pizzulo M, Reale C, Zotti T, Vito P, Stilo R - FEBS Open Bio (2015)

Bottom Line: Functionally, tBCL10 activates NF-κB transcription factor and is inhibited by the deubiquitinating enzyme A20.Finally, depletion experiments indicate that tBCL10 can functionally replace the human protein.This work demonstrates the evolutionary conservation of the mechanism of NF-κB activation through the CBM complex, and indicates that the rainbow trout O . mykiss can serve as a model organism to study this pathway.

View Article: PubMed Central - PubMed

Affiliation: Biogem, Via Camporeale, Ariano Irpino (AV), Italy.

ABSTRACT
The complexes formed by BCL10, MALT1 and members of the family of CARMA proteins have recently been the focus of much attention because they represent a key mechanism for regulating activation of the transcription factor NF-κB. Here, we report the functional characterization of a novel isoform of BCL10 in the trout Oncorhynchus mykiss, which we named tBCL10. tBCL10 dimerizes, binds to components of the CBM complex and forms cytoplasmic filaments. Functionally, tBCL10 activates NF-κB transcription factor and is inhibited by the deubiquitinating enzyme A20. Finally, depletion experiments indicate that tBCL10 can functionally replace the human protein. This work demonstrates the evolutionary conservation of the mechanism of NF-κB activation through the CBM complex, and indicates that the rainbow trout O . mykiss can serve as a model organism to study this pathway.

No MeSH data available.


Alignment and phylogenetic tree of tBCL10. (A) Alignment of tBCL10 sequence with the human BCL10 sequence and the consensus sequences generated by aligning the BCL10 sequences of Chordata and the CARD domains of three Invertebrata proteins. At the top of the alignment the six alpha helix regions of the CARD are shown. Amino acid numbering refers to the tBCL10 sequence. The alignment was using ClustalW and the printout from multiple-aligned sequences was done with BOXSHADE. The black background designates identical amino acids, the gray background conservative substitutions. Colored rectangles indicate amino acids conserved among the sequences examined. The sequences used for generation of the consensus are available in Supplementary Material. (B) Phylogenetic tree analysis of BCL10 proteins. The phylogenetic tree was constructed based on the full-length amino acid sequences using the neighbor-joining method within the Mega program. The sequences used for alignment and generation of the consensus are available in Supplementary Material. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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f0005: Alignment and phylogenetic tree of tBCL10. (A) Alignment of tBCL10 sequence with the human BCL10 sequence and the consensus sequences generated by aligning the BCL10 sequences of Chordata and the CARD domains of three Invertebrata proteins. At the top of the alignment the six alpha helix regions of the CARD are shown. Amino acid numbering refers to the tBCL10 sequence. The alignment was using ClustalW and the printout from multiple-aligned sequences was done with BOXSHADE. The black background designates identical amino acids, the gray background conservative substitutions. Colored rectangles indicate amino acids conserved among the sequences examined. The sequences used for generation of the consensus are available in Supplementary Material. (B) Phylogenetic tree analysis of BCL10 proteins. The phylogenetic tree was constructed based on the full-length amino acid sequences using the neighbor-joining method within the Mega program. The sequences used for alignment and generation of the consensus are available in Supplementary Material. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Mentions: The tBCL10 cDNA encodes for a protein of 207 amino acids, with a predicted molecular mass of 23 kDa (Table 2 and Fig. 1A). The overall amino acidic identity of tBCL10 to hBCL10 is 40% (Table 2). The major amino acidic differences between the two proteins are located at the carboxy-terminal of the polypeptides, whereas the amino-terminal CARD domains of tBCL10 (amino acids 6–116) and hBCL10 (amino acids 8–115) share 59% identity (Table 2). Sequence analysis shows that several residues that have been demonstrated to be necessary for the biological activity of hBCL10, namely R36, D39, L41, R42, E53 and G78 [4,15,20–23], are conserved in tBCL10. On the other hand, the residue S141, which is implicated in attenuation of hBCL10 signaling [24], is not conserved in tBCL10 (Fig. 1A). A phylogenetic tree was constructed by the neighbor-joining method using MEGA [14], and it shows that the sequences of rainbow trout BCL10a and BCL10b isoforms clusterize within the fish BCL10 sequences (Fig. 1B).


Functional characterization of a BCL10 isoform in the rainbow trout Oncorhynchus mykiss.

Mazzone P, Scudiero I, Coccia E, Ferravante A, Paolucci M, D'Andrea EL, Varricchio E, Pizzulo M, Reale C, Zotti T, Vito P, Stilo R - FEBS Open Bio (2015)

Alignment and phylogenetic tree of tBCL10. (A) Alignment of tBCL10 sequence with the human BCL10 sequence and the consensus sequences generated by aligning the BCL10 sequences of Chordata and the CARD domains of three Invertebrata proteins. At the top of the alignment the six alpha helix regions of the CARD are shown. Amino acid numbering refers to the tBCL10 sequence. The alignment was using ClustalW and the printout from multiple-aligned sequences was done with BOXSHADE. The black background designates identical amino acids, the gray background conservative substitutions. Colored rectangles indicate amino acids conserved among the sequences examined. The sequences used for generation of the consensus are available in Supplementary Material. (B) Phylogenetic tree analysis of BCL10 proteins. The phylogenetic tree was constructed based on the full-length amino acid sequences using the neighbor-joining method within the Mega program. The sequences used for alignment and generation of the consensus are available in Supplementary Material. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4372615&req=5

f0005: Alignment and phylogenetic tree of tBCL10. (A) Alignment of tBCL10 sequence with the human BCL10 sequence and the consensus sequences generated by aligning the BCL10 sequences of Chordata and the CARD domains of three Invertebrata proteins. At the top of the alignment the six alpha helix regions of the CARD are shown. Amino acid numbering refers to the tBCL10 sequence. The alignment was using ClustalW and the printout from multiple-aligned sequences was done with BOXSHADE. The black background designates identical amino acids, the gray background conservative substitutions. Colored rectangles indicate amino acids conserved among the sequences examined. The sequences used for generation of the consensus are available in Supplementary Material. (B) Phylogenetic tree analysis of BCL10 proteins. The phylogenetic tree was constructed based on the full-length amino acid sequences using the neighbor-joining method within the Mega program. The sequences used for alignment and generation of the consensus are available in Supplementary Material. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Mentions: The tBCL10 cDNA encodes for a protein of 207 amino acids, with a predicted molecular mass of 23 kDa (Table 2 and Fig. 1A). The overall amino acidic identity of tBCL10 to hBCL10 is 40% (Table 2). The major amino acidic differences between the two proteins are located at the carboxy-terminal of the polypeptides, whereas the amino-terminal CARD domains of tBCL10 (amino acids 6–116) and hBCL10 (amino acids 8–115) share 59% identity (Table 2). Sequence analysis shows that several residues that have been demonstrated to be necessary for the biological activity of hBCL10, namely R36, D39, L41, R42, E53 and G78 [4,15,20–23], are conserved in tBCL10. On the other hand, the residue S141, which is implicated in attenuation of hBCL10 signaling [24], is not conserved in tBCL10 (Fig. 1A). A phylogenetic tree was constructed by the neighbor-joining method using MEGA [14], and it shows that the sequences of rainbow trout BCL10a and BCL10b isoforms clusterize within the fish BCL10 sequences (Fig. 1B).

Bottom Line: Functionally, tBCL10 activates NF-κB transcription factor and is inhibited by the deubiquitinating enzyme A20.Finally, depletion experiments indicate that tBCL10 can functionally replace the human protein.This work demonstrates the evolutionary conservation of the mechanism of NF-κB activation through the CBM complex, and indicates that the rainbow trout O . mykiss can serve as a model organism to study this pathway.

View Article: PubMed Central - PubMed

Affiliation: Biogem, Via Camporeale, Ariano Irpino (AV), Italy.

ABSTRACT
The complexes formed by BCL10, MALT1 and members of the family of CARMA proteins have recently been the focus of much attention because they represent a key mechanism for regulating activation of the transcription factor NF-κB. Here, we report the functional characterization of a novel isoform of BCL10 in the trout Oncorhynchus mykiss, which we named tBCL10. tBCL10 dimerizes, binds to components of the CBM complex and forms cytoplasmic filaments. Functionally, tBCL10 activates NF-κB transcription factor and is inhibited by the deubiquitinating enzyme A20. Finally, depletion experiments indicate that tBCL10 can functionally replace the human protein. This work demonstrates the evolutionary conservation of the mechanism of NF-κB activation through the CBM complex, and indicates that the rainbow trout O . mykiss can serve as a model organism to study this pathway.

No MeSH data available.