N-Glycosylation Regulates Pannexin 2 Localization but Is Not Required for Interacting with Pannexin 1

Sanchez-Pupo, Rafael E; Johnston, Danielle; Peñuela, Silvia

doi:10.3390/ijms19071837

Cited by 19 publications

(15 citation statements)

References 43 publications

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“…We therefore investigated the localization, glycosylation, and oligomerization profiles of EGFP-tagged Panx1CT deletion mutants in HEK293T cells, which lack appreciable endogenous Panx1 expression at the protein level (however, see Sanchez-Pupo et al . 12 ).…”

Section: Introductionmentioning

confidence: 99%

A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization

Epp

Ebert

Sanchez-Arias

et al. 2019

Sci Rep

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The Pannexin 1 (Panx1) ion and metabolite channel is expressed in a wide variety of cells where it regulates a number of cell behaviours including proliferation and differentiation. Panx1 is expressed on the cell surface as well as intracellular membranes. Previous work suggests that a region within the proximal Panx1 C-terminus (Panx1CT) regulates cell surface localization. Here we report the discovery of a putative leucine-rich repeat (LRR) motif in the proximal Panx1CT necessary for Panx1 cell surface expression in HEK293T cells. Deletion of the putative LRR motif results in significant loss of Panx1 cell surface distribution. Outcomes of complementary cell surface oligomerization and glycosylation state analyses were consistent with reduced cell surface expression of Panx1 LRR deletion mutants. Of note, the oligomerization analysis revealed the presence of putative dimers and trimers of Panx1 at the cell surface. Expression of Panx1 increased HEK293T cell growth and reduced doubling time, while expression of a Panx1 LRR deletion mutant (highly conserved segment) did not reproduce this effect. In summary, here we discovered the presence of a putative LRR motif in the Panx1CT that impacts on Panx1 cell surface localization. Overall these findings provide new insights into the molecular mechanisms underlying C-terminal regulation of Panx1 trafficking and raise potential new lines of investigation with respect to Panx1 oligomerization and glycosylation.

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Section: Introductionmentioning

confidence: 99%

A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization

Epp

Ebert

Sanchez-Arias

et al. 2019

Sci Rep

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“…Due to their relatively low Panx1 endogenous expression ( Sanchez-Pupo et al. , 2018 ) and large cytoplasm to visualize PANX1 localization, normal rat kidney (NRK) cells were used to transiently overexpress the selected PANX1 variants.…”

Section: Resultsmentioning

confidence: 99%

“…Due to their relatively low Panx1 endogenous expression (Sanchez-Pupo et al, 2018) and large cytoplasm to visualize PANX1 localization, normal rat kidney (NRK) cells were used to transiently overexpress the selected PANX1 variants. Most of the variants, except for Y150F and Q264*, produced PANX1 banding patterns on immunoblots similar to Q5 (WT) consisting of Gly0, Gly1, and Gly2 (Figure 1B).…”

Section: Y150f Alters Normal Panx1 Protein Banding Pattern Indicativementioning

confidence: 99%

Pannexin 1 mutation found in melanoma tumor reduces phosphorylation, glycosylation, and trafficking of the channel-forming protein

Nouri‐Nejad

O'Donnell

Patil

et al. 2021

MBoC

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Pannexin 1 (PANX1) is a glycoprotein that forms large pore channels capable of passing ions and metabolites such as ATP for cellular communication. PANX1 has been implicated in many diseases including breast cancer and melanoma, where inhibition or deletion of PANX1 reduced the tumorigenic and metastatic properties of the cancer cells. We interrogated the effect of single amino acid changes in various PANX1 domains, using naturally occurring variants reported in cancer patient tumors. We found that a previously reported variant (Q5H), is present in cancer cells, but was not different from the wildtype (Q5) in glycosylation, trafficking, or channel function, and did not affect cellular properties. We discovered that the Q5H variant is in fact the highly conserved ancestral allele of PANX1, with 89% of humans carrying at least one Q5H allele. Another mutated form Y150F, found in a melanoma patient tumour, prevented phosphorylation at Y150 as well as complex N-glycosylation, while increasing intracellular localization. SRC is the predicted kinase to phosphorylate the Y150 residue, and its phosphorylation is not likely to be constitutive, but rather dynamically regulated. The Y150 phosphorylation site is the first one reported to play a role in regulating post-translational modifications and trafficking of PANX1, with potential consequences on its large-pore channel structure and function in melanoma cells.

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“…Consistent with this finding, vertebrate pannexin proteins have N-glycosylation sites at the extracellular loops, whereas nearly all connexins do not have the N-glycosylation sites. N-glycosylation of pannexins regulates the subcellular localization and intermixing of different pannexins for channel formation [ 49 , 50 ]. Additionally, N-glycosylation of pannexin inhibits the intercellular docking of neighboring pannexin channels and the formation of gap junctions [ 50 , 51 ].…”

Section: Resultsmentioning

confidence: 99%

Identification and classification of innexin gene transcripts in the central nervous system of the terrestrial slug Limax valentianus

et al. 2021

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Intercellular gap junction channels and single-membrane channels have been reported to regulate electrical synapse and the brain function. Innexin is known as a gap junction-related protein in invertebrates and is involved in the formation of intercellular gap junction channels and single-cell membrane channels. Multiple isoforms of innexin protein in each species enable the precise regulation of channel function. In molluscan species, sequence information of innexins is still limited and the sequences of multiple innexin isoforms have not been classified. This study examined the innexin transcripts expressed in the central nervous system of the terrestrial slug Limax valentianus and identified 16 transcripts of 12 innexin isoforms, including the splicing variants. We performed phylogenetic analysis and classified the isoforms with other molluscan innexin sequences. Next, the phosphorylation, N-glycosylation, and S-nitrosylation sites were predicted to characterize the innexin isoforms. Further, we identified 16 circular RNA sequences of nine innexin isoforms in the central nervous system of Limax. The identification and classification of molluscan innexin isoforms provided novel insights for understanding the regulatory mechanism of innexin in this phylum.

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N-Glycosylation Regulates Pannexin 2 Localization but Is Not Required for Interacting with Pannexin 1

Cited by 19 publications

References 43 publications

A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization

A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization

Pannexin 1 mutation found in melanoma tumor reduces phosphorylation, glycosylation, and trafficking of the channel-forming protein

Identification and classification of innexin gene transcripts in the central nervous system of the terrestrial slug Limax valentianus

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