TY - JOUR
T1 - Identification and functional characterization of three NoLS (nucleolar localisation signals) mutations of the CDC73 Gene
AU - Pazienza, Valerio
AU - La Torre, Annamaria
AU - Baorda, Filomena
AU - Alfarano, Michela
AU - Chetta, Massimiliano
AU - Muscarella, Lucia Anna
AU - Battista, Claudia
AU - Copetti, Massimiliano
AU - Kotzot, Dieter
AU - Kapelari, Klaus
AU - Mahmoud Yousef Alabdulrazzaq, Dalia
AU - Perlman, Kusiel
AU - Sochett, Etienne
AU - Cole, David E.C.
AU - Pellegrini, Fabio
AU - Canaff, Lucie
AU - Hendy, Geoffrey N.
AU - D'Agruma, Leonardo
AU - Zelante, Leopoldo
AU - Carella, Massimo
AU - Scillitani, Alfredo
AU - Guarnieri, Vito
PY - 2013/12/5
Y1 - 2013/12/5
N2 - Abstract Hyperparathyroidism Jaw-Tumour Syndrome (HPT-JT) is characterized by primary hyperparathyroidism (PHPT), maxillary/mandible ossifying fibromas and by parathyroid carcinoma in 15% of cases. Inactivating mutations of the tumour suppressor CDC73/HRPT2 gene have been found in HPT-JT patients and also as genetic determinants of sporadic parathyroid carcinoma/atypical adenomas and, rarely, typical adenomas, in familial PHPT. Here we report the genetic and molecular analysis of the CDC73/HRPT2 gene in three patients affected by PHPT due to atypical and typical parathyroid adenomas, in one case belonging to familial PHPT. Flag-tagged WT and mutant CDC73/ HRPT2 proteins were transiently transfected in HEK293 cells and functional assays were performed in order to investigate the effect of the variants on the whole protein expression, nuclear localization and cell overgrowth induction. We identified four CDC73/HRPT2 gene mutations, three germline (c.679-680delAG, p.Val85-Val86del and p.Glu81-Pro84del), one somatic (p.Arg77Pro). In three cases the mutation was located within the Nucleolar Localisation Signals (NoLS). The three NoLS variants led to instability either of the corresponding mutated protein or mRNA or both. When transfected in HEK293 cells, NoLS mutated proteins mislocalized with a predeliction for cytoplasmic or nucleo-cytoplasmic localization and, finally, they resulted in overgrowth, consistent with a dominant negative interfering effect in the presence of the endogenous protein.
AB - Abstract Hyperparathyroidism Jaw-Tumour Syndrome (HPT-JT) is characterized by primary hyperparathyroidism (PHPT), maxillary/mandible ossifying fibromas and by parathyroid carcinoma in 15% of cases. Inactivating mutations of the tumour suppressor CDC73/HRPT2 gene have been found in HPT-JT patients and also as genetic determinants of sporadic parathyroid carcinoma/atypical adenomas and, rarely, typical adenomas, in familial PHPT. Here we report the genetic and molecular analysis of the CDC73/HRPT2 gene in three patients affected by PHPT due to atypical and typical parathyroid adenomas, in one case belonging to familial PHPT. Flag-tagged WT and mutant CDC73/ HRPT2 proteins were transiently transfected in HEK293 cells and functional assays were performed in order to investigate the effect of the variants on the whole protein expression, nuclear localization and cell overgrowth induction. We identified four CDC73/HRPT2 gene mutations, three germline (c.679-680delAG, p.Val85-Val86del and p.Glu81-Pro84del), one somatic (p.Arg77Pro). In three cases the mutation was located within the Nucleolar Localisation Signals (NoLS). The three NoLS variants led to instability either of the corresponding mutated protein or mRNA or both. When transfected in HEK293 cells, NoLS mutated proteins mislocalized with a predeliction for cytoplasmic or nucleo-cytoplasmic localization and, finally, they resulted in overgrowth, consistent with a dominant negative interfering effect in the presence of the endogenous protein.
UR - http://www.scopus.com/inward/record.url?scp=84891910304&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0082292
DO - 10.1371/journal.pone.0082292
M3 - Article
C2 - 24340015
AN - SCOPUS:84891910304
VL - 8
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 12
M1 - e82292
ER -