Effect of mutations of murine lens alphaB crystallin on transfected neural cell viability and cellular translocation in response to stress

FEBS Lett. 1998 Oct 30;438(1-2):25-31. doi: 10.1016/s0014-5793(98)01260-5.

Abstract

We examined the influence of over-expressed native and mutant murine lens alphaB crystallin on the response of a murine neural cell line to heat and ionic strength shock. Native and mutant (F27R and KK174/175LL) murine alphaB crystallin amplicons were subcloned into a Lac-Switch IPTG-inducible RSV promoter eukaryotic vector, and transfected into N1E-115 cells using lipofectin. Expression was induced maximally 8 h after addition of IPTG (optimal final concentration 1 mM) to the medium. Cells grew normally after transfection with native and mutant murine alphaB crystallin. We demonstrated expression of the protein using specific anti-alpha crystallin antibodies. Viability under severe heat and ionic strength stress increased when cells had been transfected with native alphaB crystallin, but not with mutants F27R or KK174/175LL. Heat shock caused translocation of both native and mutant alphaB crystallins into the central region of the cells. These results show that mutations in alphaB crystallin that effect its chaperone-like activity may also influence viability of N1E-115 neural cells under stress, while not influencing the distribution of the protein within the cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Movement
  • Cell Survival
  • Crystallins / genetics*
  • Crystallins / metabolism
  • Heat-Shock Response*
  • Humans
  • Mice
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Mutation*
  • Neuroblastoma
  • Neurons / cytology*
  • Neurons / physiology
  • Osmolar Concentration
  • Phenotype
  • Sequence Alignment
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Crystallins