A differential proteomic approach to identify proteins associated with thyroid cell transformation.
Tumour suppressor p53 is a transcription factor essential for DNA damage checkpoints during cellular response to stress. Mutations in the p53 gene are the most common genetic alterations found in human tumours; most pathogenetic modifications are missense mutations that abolish the p53 DNA-binding function. In the same cell type, distinct p53 missense mutations may determine different phenotypes. The PC Cl3 cell line retains several markers of thyroid differentiation in vitro. Introduction of the V143A mutant p53 allele, which abolishes the p53 DNA-binding function, leads to loss of differentiation markers as well as TSH dependency for growth. Conversely, PC Cl3 cells transfected with the S392A mutant p53 allele, presenting the mutation located outside the DNA-binding domain, show only loss of TSH dependency for growth. To identify molecular differences existing between PC Cl3 cell lines transformed by the V143A and the S392A mutant alleles, a differential proteomic approach was used. Two-dimensional gel electrophoresis analyses indicated that expression of a significant portion of protein species was modified by both p53 mutants. In fact, compared with wild-type PC Cl3 cells, modification of expression in V143A mutant cells occurred in 23.6% of the entire protein species. Conversely, modification of S392A mutant cells affected 14.0% of total proteins. Among these components, 8.3% were common to both mutants. Several of these proteins were identified by mass spectrometry procedures; some proteins, such as HSP90 and T-complex proteins, are already known to be related to p53 function.
Paron
I
I
Dipartimento di Scienze e Tecnologie Biomediche, Università degli Studi di Udine, Udine, Italy.
D'Ambrosio
C
C
Scaloni
A
A
Berlingieri
M T
MT
Pallante
P L
PL
Fusco
A
A
Bivi
N
N
Tell
G
G
Damante
G
G
D037282:Calreticulin / Q000302:isolation & purification; D002471:Cell Transformation, Neoplastic / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D037483:Galectin 1 / Q000302:isolation & purification; D018841:HSP90 Heat-Shock Proteins / Q000302:isolation & purification; D006801:Humans; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism*; D020543:Proteome; D013961:Thyroid Gland / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014746:Vimentin / Q000302:isolation & purification
D037282:Calreticulin; D037483:Galectin 1; D018841:HSP90 Heat-Shock Proteins; D009363:Neoplasm Proteins; D020543:Proteome; D016159:Tumor Suppressor Protein p53; D014746:Vimentin