Pathogenic cysteine mutations affect progranulin function and production of mature granulins

J Neurochem. 2010 Mar;112(5):1305-15. doi: 10.1111/j.1471-4159.2009.06546.x. Epub 2009 Dec 17.

Abstract

Frontotemporal dementia with ubiquitin-positive inclusions (FTLD-U) can be caused by mutations in the progranulin gene (GRN). Progranulin (PGRN) is a cysteine-rich growth factor, which is proteolytically cleaved by elastase to produce several granulins (GRNs). All FTLD-U mutations in GRN characterized to date result in reduced secreted PGRN protein. We recently reported a Spanish family with progressive non-fluent aphasia and dementia in which a novel C521Y mutation segregates with disease. A second cysteine mutation (C139R) has also been reported to be disease specific. Allele-specific mRNA expression assays in brain reveal that the C521Y mutant allele is expressed at similar levels to the wild-type allele. Furthermore, plasma PGRN levels in C521Y carriers are comparable with non-carrier family relatives, suggesting that the mutation does not affect PGRN protein expression and secretion in vivo. Despite normal PGRN levels C521Y and C139R mutant GRNs show reduced neurite growth-stimulating activity in vitro. Further study revealed that these mutations also cause impaired cleavage of PGRN by elastase. Our data suggest that these mutations affect the function of full-length PGRN as well as elastase cleavage of PGRN into GRNs, leading to neurodegeneration.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arginine / genetics
  • Axons / physiology
  • Brain / cytology
  • Brain / pathology
  • Cell Enlargement
  • Cell Line, Transformed
  • Cysteine / genetics*
  • Dithiothreitol / pharmacology
  • Embryo, Mammalian
  • Enzyme-Linked Immunosorbent Assay / methods
  • Family Health
  • Frontotemporal Dementia / genetics*
  • Frontotemporal Dementia / pathology
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / genetics
  • Genetic Predisposition to Disease*
  • Green Fluorescent Proteins / genetics
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Intercellular Signaling Peptides and Proteins / physiology*
  • Motor Neurons / cytology
  • Motor Neurons / drug effects
  • Motor Neurons / metabolism
  • Mutation / genetics*
  • Neurites
  • Pancreatic Elastase / pharmacology
  • Progranulins
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Wistar
  • Transfection
  • Tyrosine / genetics

Substances

  • GRN protein, human
  • Intercellular Signaling Peptides and Proteins
  • Progranulins
  • RNA, Messenger
  • Green Fluorescent Proteins
  • Tyrosine
  • Arginine
  • Pancreatic Elastase
  • Cysteine
  • Dithiothreitol