BAC-mediated transgenic expression of fluorescent autophagic protein Beclin 1 reveals a role for Beclin 1 in lymphocyte development

Cell Death Differ. 2008 Sep;15(9):1385-95. doi: 10.1038/cdd.2008.59. Epub 2008 May 2.

Abstract

Beclin 1/Atg6 is an essential component of the evolutionary conserved PtdIns(3)-kinase (Vps34) protein complex that regulates macroautophagy (autophagy) in eukaryotic cells and also interacts with antiapoptotic Bcl-2 family members, Bcl-2, and Bcl-x(L). To elucidate the physiological function of Beclin 1, we generated transgenic mice producing a green fluorescent Beclin 1 protein (Beclin 1-GFP) under Beclin 1 endogenous regulation. The beclin 1-GFP transgene is functional because it completely rescues early embryonic lethality in beclin 1-deficient mice. The transgenic mice appear normal, with undetected change in basal autophagy levels in different tissues, despite the additional expression of functional Beclin 1-GFP. Staining of Beclin 1-GFP shows mostly diffuse cytoplasmic distribution in various tissues. Detailed analysis of the transgene expression by flow cytometry reveals a Bcl-2-like biphasic expression pattern in developing T and B cells, as well as differential regulation of expression in mature versus immature thymocytes following in vitro stimulation. Moreover, thymocytes expressing high Beclin 1-GFP levels appear increasingly sensitive to glucocorticoid-induced apoptosis in vitro. Our results, therefore, support a role for Beclin 1 in lymphocyte development involving cross talk between autophagy and apoptosis.

Publication types

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

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism
  • Apoptosis Regulatory Proteins / physiology*
  • Apoptosis*
  • Autophagy*
  • B-Lymphocytes / immunology
  • Beclin-1
  • CD4-Positive T-Lymphocytes / immunology
  • Chromosomes, Artificial, Bacterial
  • Dexamethasone / pharmacology
  • Fluorescent Dyes / analysis
  • Glucocorticoids / pharmacology
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • Lymphocyte Activation
  • Mice
  • Mice, Transgenic
  • Proteins / genetics
  • Proteins / metabolism
  • Proteins / physiology*
  • T-Lymphocytes / immunology*
  • Thymus Gland / cytology
  • Thymus Gland / immunology

Substances

  • Apoptosis Regulatory Proteins
  • Beclin-1
  • Becn1 protein, mouse
  • Fluorescent Dyes
  • Glucocorticoids
  • Proteins
  • Green Fluorescent Proteins
  • Dexamethasone