Suppression of proliferative defects associated with processing-defective lamin A mutants by hTERT or inactivation of p53.

Hutchinson-Gilford progeria syndrome (HGPS) is a rare, debilitating disease with early mortality and rapid onset of aging-associated pathologies. It is linked to mutations in LMNA, which encodes A-type nuclear lamins. The most frequent HGPS-associated LMNA mutation results in a protein, termed progerin, with an internal 50 amino acid deletion and, unlike normal A-type lamins, stable farnesylation. The cellular consequences of progerin expression underlying the HGPS phenotype remain poorly understood. Here, we stably expressed lamin A mutants, including progerin, in otherwise identical primary human fibroblasts to compare the effects of different mutants on nuclear morphology and cell proliferation. We find that expression of progerin leads to inhibition of proliferation in a high percentage of cells and slightly premature senescence in the population. Expression of a stably farnesylated mutant of lamin A phenocopied the immediate proliferative defects but did not result in premature senescence. Either p53 inhibition or, more surprisingly, expression of the catalytic subunit of telomerase (hTERT) suppressed the early proliferative defects associated with progerin expression. These findings lead us to propose that progerin may interfere with telomere structure or metabolism in a manner suppressible by increased telomerase levels and possibly link mechanisms leading to progeroid phenotypes to those of cell immortalization.

[1]  Richard T. Lee,et al.  Increased mechanosensitivity and nuclear stiffness in Hutchinson–Gilford progeria cells: effects of farnesyltransferase inhibitors , 2008, Aging cell.

[2]  Kaushik Sengupta,et al.  Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. , 2008, Genes & development.

[3]  T. Misteli,et al.  Lamin A-dependent misregulation of adult stem cells associated with accelerated ageing , 2008, Nature Cell Biology.

[4]  P. Rabinovitch,et al.  Accelerated telomere shortening and replicative senescence in human fibroblasts overexpressing mutant and wild-type lamin A. , 2008, Experimental cell research.

[5]  Sandy S. Tungteakkhun,et al.  Cellular binding partners of the human papillomavirus E6 protein , 2008, Archives of Virology.

[6]  B. Kennedy,et al.  Evidence that Proteasome-Dependent Degradation of the Retinoblastoma Protein in Cells Lacking A-Type Lamins Occurs Independently of Gankyrin and MDM2 , 2007, PloS one.

[7]  N. Lévy,et al.  An association of Hutchinson–Gilford progeria and malignancy , 2007, American journal of medical genetics. Part A.

[8]  B. Kennedy,et al.  Werner and Hutchinson–Gilford progeria syndromes: mechanistic basis of human progeroid diseases , 2007, Nature Reviews Molecular Cell Biology.

[9]  G. Sluder,et al.  p53-independent abrogation of a postmitotic checkpoint contributes to human papillomavirus E6-induced polyploidy. , 2007, Cancer research.

[10]  R. Foisner,et al.  Nucleoplasmic lamins and their interaction partners, LAP2α, Rb, and BAF, in transcriptional regulation , 2007, The FEBS journal.

[11]  Richard T. Lee,et al.  Lamins A and C but Not Lamin B1 Regulate Nuclear Mechanics* , 2006, Journal of Biological Chemistry.

[12]  Y. Gruenbaum,et al.  The nuclear lamina and its proposed roles in tumorigenesis: projection on the hematologic malignancies and future targeted therapy. , 2006, Journal of structural biology.

[13]  M. Bergo,et al.  A farnesyltransferase inhibitor improves disease phenotypes in mice with a Hutchinson-Gilford progeria syndrome mutation. , 2006, The Journal of clinical investigation.

[14]  B. Kennedy,et al.  Stabilization of the Retinoblastoma Protein by A-Type Nuclear Lamins Is Required for INK4A-Mediated Cell Cycle Arrest , 2006, Molecular and Cellular Biology.

[15]  Tom Misteli,et al.  Distinct structural and mechanical properties of the nuclear lamina in Hutchinson–Gilford progeria syndrome , 2006, Proceedings of the National Academy of Sciences.

[16]  A. Klingelhutz,et al.  Human Papillomavirus Type 16 E6 Activates NF-κB, Induces cIAP-2 Expression, and Protects against Apoptosis in a PDZ Binding Motif-Dependent Manner , 2006, Journal of Virology.

[17]  T. Misteli,et al.  Lamin A-Dependent Nuclear Defects in Human Aging , 2006, Science.

[18]  Stephen G Young,et al.  A Protein Farnesyltransferase Inhibitor Ameliorates Disease in a Mouse Model of Progeria , 2006, Science.

[19]  Richard L. Frock,et al.  Lamin A/C and emerin are critical for skeletal muscle satellite cell differentiation. , 2006, Genes & development.

[20]  J. O'connor,et al.  A mechanism of AP-1 suppression through interaction of c-Fos with lamin A/C. , 2006, Genes & development.

[21]  M. W. Glynn,et al.  Incomplete processing of mutant lamin A in Hutchinson-Gilford progeria leads to nuclear abnormalities, which are reversed by farnesyltransferase inhibition. , 2005, Human molecular genetics.

[22]  B. Kennedy,et al.  HIV protease inhibitors block adipocyte differentiation independently of lamin A/C , 2005, AIDS.

[23]  Joel H. Janes,et al.  Correction of cellular phenotypes of Hutchinson-Gilford Progeria cells by RNA interference , 2005, Human Genetics.

[24]  M. Gelb,et al.  Inhibiting farnesylation reverses the nuclear morphology defect in a HeLa cell model for Hutchinson-Gilford progeria syndrome. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Ignacio Varela,et al.  Accelerated ageing in mice deficient in Zmpste24 protease is linked to p53 signalling activation , 2005, Nature.

[26]  Karen N Conneely,et al.  Inhibiting farnesylation of progerin prevents the characteristic nuclear blebbing of Hutchinson-Gilford progeria syndrome. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Gelb,et al.  Blocking protein farnesyltransferase improves nuclear shape in fibroblasts from humans with progeroid syndromes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[28]  E. Parry,et al.  Molecular cytogenetic insights into the ageing syndrome Hutchinson-Gilford Progeria (HGPS) , 2005, Cytogenetic and Genome Research.

[29]  M. Gelb,et al.  Blocking protein farnesyltransferase improves nuclear blebbing in mouse fibroblasts with a targeted Hutchinson-Gilford progeria syndrome mutation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[30]  David J. Chen,et al.  Genomic instability in laminopathy-based premature aging , 2005, Nature Medicine.

[31]  Richard L. Frock,et al.  A-type nuclear lamins, progerias and other degenerative disorders , 2005, Mechanisms of Ageing and Development.

[32]  T. Misteli,et al.  Reversal of the cellular phenotype in the premature aging disease Hutchinson-Gilford progeria syndrome , 2005, Nature Medicine.

[33]  E. Furth,et al.  Telomere Shortening Exposes Functions for the Mouse Werner and Bloom Syndrome Genes , 2004, Molecular and Cellular Biology.

[34]  R. DePinho,et al.  Essential role of limiting telomeres in the pathogenesis of Werner syndrome , 2004, Nature Genetics.

[35]  Richard L. Frock,et al.  A-type lamins regulate retinoblastoma protein function by promoting subnuclear localization and preventing proteasomal degradation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[36]  R. Faragher,et al.  Fibroblast clones from patients with Hutchinson–Gilford progeria can senesce despite the presence of telomerase , 2004, Experimental Gerontology.

[37]  Richard T. Lee,et al.  Lamin A/C deficiency causes defective nuclear mechanics and mechanotransduction. , 2004, The Journal of clinical investigation.

[38]  Pierre Cau,et al.  Lamin A Truncation in Hutchinson-Gilford Progeria , 2003, Science.

[39]  L. Mounkes,et al.  A progeroid syndrome in mice is caused by defects in A-type lamins , 2003, Nature.

[40]  Laura Scott,et al.  Recurrent de novo point mutations in lamin A cause Hutchinson–Gilford progeria syndrome , 2003, Nature.

[41]  R. Hegele,et al.  LMNA is mutated in Hutchinson-Gilford progeria (MIM 176670) but not in Wiedemann-Rautenstrauch progeroid syndrome (MIM 264090) , 2003, Journal of Human Genetics.

[42]  Karl Münger,et al.  Human papillomavirus immortalization and transformation functions. , 2002, Virus research.

[43]  J. Campisi,et al.  Reversible Manipulation of Telomerase Expression and Telomere Length , 2002, The Journal of Biological Chemistry.

[44]  Sui Huang,et al.  Alteration of nuclear lamin organization inhibits RNA polymerase II–dependent transcription , 2002, The Journal of cell biology.

[45]  D. Galloway,et al.  E Box-Dependent Activation of Telomerase by Human Papillomavirus Type 16 E6 Does Not Require Induction of c-myc , 2001, Journal of Virology.

[46]  C. Woodworth,et al.  Papillomavirus Type 16 Oncogenes Downregulate Expression of Interferon-Responsive Genes and Upregulate Proliferation-Associated and NF-κB-Responsive Genes in Cervical Keratinocytes , 2001, Journal of Virology.

[47]  J. Shay,et al.  The establishment of telomerase-immortalized cell lines representing human chromosome instability syndromes. , 2000, Human molecular genetics.

[48]  R. Faragher,et al.  Telomerase prevents the accelerated cell ageing of Werner syndrome fibroblasts , 2000, Nature Genetics.

[49]  R. Goldman,et al.  Disruption of Nuclear Lamin Organization Alters the Distribution of Replication Factors and Inhibits DNA Synthesis , 1997, The Journal of cell biology.

[50]  J. McDougall,et al.  Telomerase activation by the E6 gene product of human papillomavirus type 16 , 1996, Nature.

[51]  C Roskelley,et al.  A biomarker that identifies senescent human cells in culture and in aging skin in vivo. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[52]  K. Ishizaki,et al.  Mutation spectrum of the retinoblastoma gene in osteosarcomas. , 1994, Cancer research.

[53]  A. Kupfer,et al.  The processing pathway of prelamin A. , 1994, Journal of cell science.

[54]  L. Wenger,et al.  Nucleoplasmic localization of prelamin A: implications for prenylation-dependent lamin A assembly into the nuclear lamina. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[55]  J. Shay,et al.  A role for both RB and p53 in the regulation of human cellular senescence. , 1991, Experimental cell research.

[56]  K. Weber,et al.  Differential timing of nuclear lamin A/C expression in the various organs of the mouse embryo and the young animal: a developmental study. , 1989, Development.

[57]  M. Kirschner,et al.  Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins , 1986, Nature.

[58]  J. Campbell,et al.  Osteosarcoma in a patient with Hutchinson-Gilford progeria. , 1978, Journal of medical genetics.