CNR - Institute of Neuroscience CNR
Institute of Neuroscience
 

Project

Structural and functional implication of protein kinase CK2 in cystic fibrosis pathogenesis

Background

The deletion of residue phenylalanine-508 (deltaF508) within NBD1 domain of cystic fibrosis transmembrane conductance regulator (CFTR) in the close proximity of a CK2 consensus sequence is the commonest mutation in cystic fibrosis (CF). Several observations prompted us to undertake a study on the structural and functional implication of CK2 in CF pathogenesis: i) NBD1 domain of wild type CFTR is phosphorylated by CK2 holoenzyme with an efficiency which is decreased by the ΔF508 mutation; ii) In epithelial cells expressing wt CFTR CK2α colocalizes to the apical membrane whereas in cell expressing ΔF508 CFTR the intracellular distribution of CK2α is perturbed; iii) The functionality of CFTR is impaired by the specific CK2 inhibitor TBB in a manner which is reversed by co-expression of a CK2 mutant refractory to inhibition; iv) Several partners of the regulatory Β-subunit of CK2 identified by an MS/affinity approach are functionally related to CFTR.

Objectives

The aim of this project is to disclose functional links between the highly pleiotropic protein kinase CK2, and cystic fibrosis (CF) pathogenesis with special reference to regulation/deregulation of cystic fibrosis transmembrane conductance regulator (CFTR) whose deletion of residue phenylalanine-508 (deltaF508) is the commonest mutation in CF.

Methods

Wild type and mutated NBD1 proteins will be used in in vitro phosphorylation assays by either the catalytic subunit or the CK2 holoenzyme. A series of NBD1-derived synthetic peptides will be used either as CK2 potential phosphoacceptor substrates or regulators.

Publications

  • Pagano MA, Arrigoni G, Marin O, Sarno S, Meggio F, Treharne KJ, Mehta A, Pinna LA (2008) Modulation of protein kinase CK2 activity by fragments of CFTR encompassing F508 may reflect functional links with cystic fibrosis pathogenesis. Biochemistry 47:7925-36.

Grants

Wellcome Trust Foundation, UK

FFC, Italy

Collaborations

  • Prof. A. Mehta, Division of Medical Sciences, Centre for Cardiovascular and Lung Biology, University of Dundee.

 

PI photo

Lorenzo Pinna

Contact information

email  E-mail

email  049 8276108

Participating staff

Mario A. Pagano
Flavio Meggio
Giorgio Cozza
Stefania Sarno
Oriano Marin