Identifying functional partners for protein/protein interactions can be a difficult challenge. We proposed the use of the operon structure of the Caenorhabditis elegans genome as a "new gene-finding tool" (Eichmuller, S., Vezzoli, V., Bazzini, C., Ritter, M., Furst, J., Jakab, M., Ravasio, A., Chwatal, S., Dossena, S., Botta, G., Meyer, G., Maier, B., Valenti, G., Lang, F., and Paulmichl, M. (2004) J. Biol. Chem. 279, 7136 7146) that could be functionally translated to the human system. Here we show the validity of this approach by studying the predicted functional interaction between ICln and HSPC038. In C. elegans, the gene encoding for the ICln homolog (icln-1) is embedded in an operon with two other genes, Nx (the human homolog of Nx is HSPC038) and Ny. ICln is a highly conserved, ubiquitously expressed multifunctional protein that plays a critical role in the regulatory volume decrease after cell swelling. Following hypotonic stress, ICln translocates from the cytosol to the plasma membrane, where it has been proposed to participate in the activation of the swelling-induced chloride current (ICl(swell)). Here we show that the interaction between human ICln and HSPC038 plays a role in volume regulation after cell swelling and that HSPC038 acts as an escort, directing ICln to the cell membrane after cell swelling and facilitating the activation of ICl(swell). Assessment of the NMR structure of HSPC038 showed the presence of a zinc finger motif. Moreover, NMR and additional biochemical techniques enabled us to identify the putative ICln/HSPC038 interacting sites, thereby explaining the functional interaction of both proteins on a molecular level.

The molecular and functional interaction between ICln and HSPC038 proteins modulates the regulation of cell volume / S. Dossena, R. Gandini, G. Tamma, V. Vezzoli, C. Nofziger, M. Tamplenizza, E. Salvioni, E. Bernardinelli, G. Meyer, G. Valenti, M. Wolf-Watz, J. Fürst, M. Paulmichl. - In: THE JOURNAL OF BIOLOGICAL CHEMISTRY. - ISSN 0021-9258. - 286:47(2011 Nov), pp. 40659-40670.

The molecular and functional interaction between ICln and HSPC038 proteins modulates the regulation of cell volume

V. Vezzoli;E. Bernardinelli;G. Meyer;
2011

Abstract

Identifying functional partners for protein/protein interactions can be a difficult challenge. We proposed the use of the operon structure of the Caenorhabditis elegans genome as a "new gene-finding tool" (Eichmuller, S., Vezzoli, V., Bazzini, C., Ritter, M., Furst, J., Jakab, M., Ravasio, A., Chwatal, S., Dossena, S., Botta, G., Meyer, G., Maier, B., Valenti, G., Lang, F., and Paulmichl, M. (2004) J. Biol. Chem. 279, 7136 7146) that could be functionally translated to the human system. Here we show the validity of this approach by studying the predicted functional interaction between ICln and HSPC038. In C. elegans, the gene encoding for the ICln homolog (icln-1) is embedded in an operon with two other genes, Nx (the human homolog of Nx is HSPC038) and Ny. ICln is a highly conserved, ubiquitously expressed multifunctional protein that plays a critical role in the regulatory volume decrease after cell swelling. Following hypotonic stress, ICln translocates from the cytosol to the plasma membrane, where it has been proposed to participate in the activation of the swelling-induced chloride current (ICl(swell)). Here we show that the interaction between human ICln and HSPC038 plays a role in volume regulation after cell swelling and that HSPC038 acts as an escort, directing ICln to the cell membrane after cell swelling and facilitating the activation of ICl(swell). Assessment of the NMR structure of HSPC038 showed the presence of a zinc finger motif. Moreover, NMR and additional biochemical techniques enabled us to identify the putative ICln/HSPC038 interacting sites, thereby explaining the functional interaction of both proteins on a molecular level.
caenorhabditis-elegans operons; membrane transposition; chloride channels; anion channel; I-CLN; translocation; activatin; cytosol; PICLN; conductance
Settore BIO/09 - Fisiologia
Settore BIO/18 - Genetica
nov-2011
Article (author)
File in questo prodotto:
File Dimensione Formato  
JBC 2011 Dossena.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 5.36 MB
Formato Adobe PDF
5.36 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/221524
Citazioni
  • ???jsp.display-item.citation.pmc??? 7
  • Scopus 18
  • ???jsp.display-item.citation.isi??? 18
social impact