Identification and Functional Characterization of Three NoLS (Nucleolar Localisation Signals) Mutations of the CDC73 Gene

Pazienza, Valerio; Torre, Annamaria la; Baorda, Filomena; Alfarano, Michela; Chetta, Massimiliano; Muscarella, Lucia Anna; Battista, Claudia; Copetti, Massimiliano; Kotzot, Dieter; Kapelari, Klaus; Al‐Abdulrazzaq, Dalia; Perlman, Kusiel; Sochett, Etienne; Cole, David E. C.; Pellegrini, Fabio; Canaff, Lucie; Hendy, Geoffrey N.; D’Agruma, Leonardo; Zelante, Leopoldo; Carella, Massimo; Scillitani, Alfredo; Guarnieri, Vito

doi:10.1371/journal.pone.0082292

Cited by 18 publications

(8 citation statements)

References 33 publications

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“…The effect of wild-type and mutant parafibromin on cell cycle and cell proliferation was also assessed and, as expected for a tumor suppressor gene, an antiproliferative activity of wild-type parafibromin was demonstrated. Importantly, this activity was not retained by the Ile60Asn mutant, which stimulated cell proliferation, thus confirming previous findings about a dominant-interfering activity of other parafibromin mutants [17] – [18] . In addition, Ile60Asn mutant parafibromin lost the ability to down-regulate c-myc expression.…”

Section: Discussionsupporting

confidence: 88%

“…Importantly, this activity was not retained by the Ile60Asn mutant, which stimulated cell proliferation, thus confirming previous findings about a dominant-interfering activity of other parafibromin mutants [17]–[18]. In addition, Ile60Asn mutant parafibromin lost the ability to down-regulate c-myc expression.…”

Section: Discussionsupporting

confidence: 86%

“…The N-terminal domain is probably crucial for parafibromin activity and has been involved by most missense mutations of the CDC73 gene [1] , [11] . Among the reported missense mutations in the N-terminal domain of parafibromin, Leu64Pro impairs parafibromin anti-proliferative activity and ability to repress cyclin D1 expression [4] ; Leu95Pro abolishes nucleolar localization and provides dominant interfering properties to parafibromin [17] ; Arg77Pro causes cytoplasm retention, decreased half-life and cell overgrowth, even in the presence of endogenous parafibromin [18] . To the best of our knowledge, no other CDC73 missense substitutions have been characterized so far.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of a New CDC73 Missense Mutation that Impairs Parafibromin Expression and Nucleolar Localization

et al. 2014

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Mutations of the Cell Division Cycle 73 (CDC73) tumor suppressor gene (previously known as HRPT2), encoding for parafibromin, are associated with the Hyperparathyroidism-Jaw Tumor (HPT-JT) syndrome, an autosomal dominant disease whose clinical manifestations are mainly parathyroid tumors and, less frequently, ossifying fibromas of the jaws, uterine and renal tumors. Most mutations of CDC73 are nonsense or frameshift, while missense mutations are rare and generally affect the N-terminal domain of parafibromin, a region that is still poorly characterized. The aim of this study was to characterize a novel somatic CDC73 missense mutation (Ile60Asn) identified in the mandibular tumor of a HPT-JT patient carrying a germline CDC73 inactivating mutation. Immunostaining of the tumor showed reduced nuclear parafibromin immunoreactivity. Western blotting and confocal microscopy of transfected cells demonstrated that the Ile60Asn mutant parafibromin was less expressed than the wild-type protein and exhibited impaired nucleolar localization. Treatment of transfected cells with translation and proteasome inhibitors demonstrated a decreased stability of the Ile60An mutant, partially due to an increase in proteasomal degradation. Overexpression of the Ile60Asn mutant led to increased cell proliferation and to accumulation in the G2/M phase of cell cycle. Moreover, mutant parafibromin lost the ability to down-regulate c-myc expression. In conclusion, our study shows that a missense mutation in the N-terminus of parafibromin, identified in an ossifying fibroma from a HPT-JT patient, stimulated cell proliferation and impaired parafibromin expression and nucleolar localization, suggesting a relevant role of the N-terminal domain for parafibromin function.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

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Characterization of a New CDC73 Missense Mutation that Impairs Parafibromin Expression and Nucleolar Localization

et al. 2014

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“…The 184 bp 5’UTR sequence of the mRNA (encoded by the CDC73 gene) was PCR amplified from normal human DNA with the forward primer having an EcoRI and the reverse primer having an SgfI restriction site. The PCR product, after digestion with these enzymes, was cloned (upstream of the open reading frame) into the EcoRI/SgfI digested pCMV6 vector that expresses the human CDC73 cDNA encoding parafibromin Myc/Flag-tagged at its COOH-terminus (Origene RC209479) [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

Large intragenic deletion of CDC73 (exons 4–10) in a three-generation hyperparathyroidism-jaw tumor (HPT-JT) syndrome family

et al. 2017

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BackgroundInactivating mutations of CDC73 cause Hyperparathyroidism-Jaw Tumour syndrome (HPT-JT), Familial Isolated Hyperparathyroidism (FIHP) and sporadic parathyroid carcinoma. We conducted CDC73 mutation analysis in an HPT-JT family and confirm carrier status of the proband’s daughter.MethodsThe proband had primary hyperparathyroidism (parathyroid carcinoma) and uterine leiomyomata. Her father and daughter had hyperparathyroidism (parathyroid adenoma) but no other manifestations of HPT-JT. CDC73 mutation analysis (sequencing of all 17 exons) and whole-genome copy number variation (CNV) analysis was done on leukocyte DNA of the three affecteds as well as the proband’s unaffected sister.ResultsA novel deletion of exons 4 to 10 of CDC73 was detected by CNV analysis in the three affecteds. A novel insertion in the 5’UTR (c.-4_-11insG) that co-segregated with the deletion was identified. By in vitro assay the 5’UTR insertion was shown to significantly impair the expression of the parafibromin protein. Screening for the mutated CDC73 confirmed carrier status in the proband’s daughter and the biochemistry and ultrasonography led to pre-emptive surgery and resolution of the hyperparathyroidism.ConclusionsA novel gross deletion mutation in CDC73 was identified in a three-generation HPT-JT family emphasizing the importance of including screening for large deletions in the molecular diagnostic protocol.Electronic supplementary materialThe online version of this article (doi:10.1186/s12881-017-0445-0) contains supplementary material, which is available to authorized users.

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“…Diagnosis of HPT-JT is important because of its genetic involvement and 24% malignant transformation. [9, 10] The nuclear medicine imaging - especially the scintigraphy parathyroid with 99m Tc-MIBI (methoxyisobutyl-isonitrile)-has an important role in outlining the diagnosis. [11] It has been estimated that approximately 70% of patients affected by this mutation may develop PHPT.…”

Section: Introductionmentioning

confidence: 99%

In silicoanalysis ofCDC73gene revealing 11 novel SNPs associated with Jaw Tumor Syndrome

Abdelmoneim¹,

Mohammed

Gadim³

et al. 2019

Preprint

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Back ground: hyperparathyroidism-jaw tumor (HPT-JT) is an autosomal dominant disorder with variable expression, with an estimated prevalence of 6.7 per 1,000 population. Genetic testing for predisposing CDC73 (HRPT2) mutations has been an important clinical advance, aimed at early detection and/or treatment to prevent advanced disease. The aim of this study is to assess the effect of SNPs on CDC73 structure and function using different bioinformatics tools. Method: Computational analysis using eight different in-silico tools including SIFT, PROVEAN, PolyPhen-2, SNAP2, PhD-SNP, SNPs&GO, PMut and Imutant were used to identify the impact on the structure and/or function of CDC73 gene that might be causing jaw tumour. Results: From (733) SNPs identified in the CDC73 gene we found that only Eleven were deleterious to the function and structure of protein and expected to cause syndrome.Conclusion: Eleven substantial genetic/molecular aberrations in CDC73 gene were identified that could serve as actionable targets for chemotherapeutic intervention in patients whose disease is no longer surgically curable.

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Identification and Functional Characterization of Three NoLS (Nucleolar Localisation Signals) Mutations of the CDC73 Gene

Cited by 18 publications

References 33 publications

Characterization of a New CDC73 Missense Mutation that Impairs Parafibromin Expression and Nucleolar Localization

Characterization of a New CDC73 Missense Mutation that Impairs Parafibromin Expression and Nucleolar Localization

Large intragenic deletion of CDC73 (exons 4–10) in a three-generation hyperparathyroidism-jaw tumor (HPT-JT) syndrome family

In silicoanalysis ofCDC73gene revealing 11 novel SNPs associated with Jaw Tumor Syndrome

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