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Pannexin 1 Ohnologs in the Teleost Lineage

Abstract

Advances in genomic analysis indicate that the early chordate lineage underwent two whole-genome duplication events in fairly rapid succession around 400–600 million years ago, and that a third duplication event punctuated the radiation of ray-finned fishes (teleosts) around 320–350 million years ago. Connexin ohnologs have been disproportionately well maintained in the teleost genome following this third event, implying that gap junction proteins are amenable to neofunctionalization. A second family of gap junction–like proteins, the pannexins, is also present in chordates, but expansion of this family following the teleost whole-genome duplication has not been addressed in the literature. In the current study we report that ohnologs of panx1 are expressed by zebrafish, and orthologs of these two genes can be found in various other teleost species. The genomic locality of each gene is described, along with sequence alignments that reveal conservation of classic pannexin-specific features/motifs. The transcripts were then cloned from cDNA for in vitro analysis, and both are shown to traffic to the plasma membrane when exogenously expressed. Furthermore, electrophysiological recordings show differences in the biophysical properties between the channels formed by these two proteins. Our results indicate that both copies of the ancestral teleost panx1 gene were conserved following the last whole-genome duplication event and, following conventional zebrafish nomenclature, should now be referred to as panx1a and panx1b.

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Acknowledgement

We thank Dr. Patricia Schulte for her kind assistance with obtaining zebrafish tissues and for critical review of the manuscript.

Author information

Correspondence to Stephen R. Bond or Christian C. Naus.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 Panx1a 3D rotation (MPG 476 kb)

Supplementary material 2 Panx1a time-lapse (MPG 1214 kb)

Supplementary material 3 Panx1b 3D rotation (MPG 352 kb)

Supplementary material 4 Panx1b time-lapse (MPG 1120 kb)

Supplementary material 5 Panx2 3D rotation (MPG 724 kb)

Supplementary material 6 Panx2 time-lapse (MPG 76 kb)

Supplementary material 7 Panx3 3D rotation (MPG 322 kb)

Supplementary material 8 Panx3 time-lapse (MPG 874 kb)

Supplementary material 1 Panx1a 3D rotation (MPG 476 kb)

Supplementary material 2 Panx1a time-lapse (MPG 1214 kb)

Supplementary material 3 Panx1b 3D rotation (MPG 352 kb)

Supplementary material 4 Panx1b time-lapse (MPG 1120 kb)

Supplementary material 5 Panx2 3D rotation (MPG 724 kb)

Supplementary material 6 Panx2 time-lapse (MPG 76 kb)

Supplementary material 7 Panx3 3D rotation (MPG 322 kb)

Supplementary material 8 Panx3 time-lapse (MPG 874 kb)

Supplementary material 9 Fig. S1 Multiple pairwise alignment of Panx1 sequences. Classic pannexin/innexin-specific residues are identified, including four cysteine residues in the extracellular loops (C), a P-X-X-X-W motif in the second transmembrane domain and a positively charged residue thought to facilitate ATP gating of the channel (⊕). A cysteine residue in Panx1a (*) has also been reported to facilitate channel activity in zebrafish but is not present in any other sequence included in this study. Predicted transmembrane domains are denoted by filled boxes (PDF 406 kb)

Fig. S2 Immunoblot confirming exogenous expression of EGFP-tagged zebrafish pannexins in HeLa cells, using an anti-GFP antibody (PDF 40 kb)

Supplementary material 11 (PDF 58 kb)

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Bond, S.R., Wang, N., Leybaert, L. et al. Pannexin 1 Ohnologs in the Teleost Lineage. J Membrane Biol 245, 483–493 (2012). https://doi.org/10.1007/s00232-012-9497-4

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Keywords

  • Pannexin
  • Teleost
  • R3 whole-genome duplication
  • Ohnolog
  • Neofunctionalization