Identification and functional characterization of three NoLS (nucleolar localisation signals) mutations of the CDC73 Gene

Valerio Pazienza, Annamaria La Torre, Filomena Baorda, Michela Alfarano, Massimiliano Chetta, Lucia Anna Muscarella, Claudia Battista, Massimiliano Copetti, Dieter Kotzot, Klaus Kapelari, Dalia Mahmoud Yousef Alabdulrazzaq, Kusiel Perlman, Etienne Sochett, David E.C. Cole, Fabio Pellegrini, Lucie Canaff, Geoffrey N. Hendy, Leonardo D'Agruma, Leopoldo Zelante, Massimo CarellaAlfredo Scillitani, Vito Guarnieri

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article numbere82292
JournalPloS one
Volume8
Issue number12
DOIs
Publication statusPublished - 5 Dec 2013

Fingerprint Dive into the research topics of 'Identification and functional characterization of three NoLS (nucleolar localisation signals) mutations of the CDC73 Gene'. Together they form a unique fingerprint.

  • Cite this

    Pazienza, V., La Torre, A., Baorda, F., Alfarano, M., Chetta, M., Muscarella, L. A., Battista, C., Copetti, M., Kotzot, D., Kapelari, K., Mahmoud Yousef Alabdulrazzaq, D., Perlman, K., Sochett, E., Cole, D. E. C., Pellegrini, F., Canaff, L., Hendy, G. N., D'Agruma, L., Zelante, L., ... Guarnieri, V. (2013). Identification and functional characterization of three NoLS (nucleolar localisation signals) mutations of the CDC73 Gene. PloS one, 8(12), [e82292]. https://doi.org/10.1371/journal.pone.0082292