Bcl-2 expressed using a retroviral vector is localized primarily in the nuclear membrane and the endoplasmic reticulum of chicken embryo fibroblasts.
A complementary DNA for human bcl-2 was cloned into the replication competent avian retrovirus vector RCASBP, and the resulting virus was used to express human Bcl-2 protein at high levels in chicken embryo fibroblasts. The expression of Bcl-2 did not transform or significantly alter the longevity of the chicken embryo fibroblasts in the presence of normal amounts of serum. However, the expression of Bcl-2 blocked c-Myc-induced apoptosis in these cells. Fractionation of the infected chicken embryo fibroblasts indicated that the protein was distributed equally between nuclear and high density cytoplasmic membranes. Immunofluorescence analysis by confocal microscopy and immunoelectron microscopy showed that the Bcl-2 protein was primarily associated with the nuclear membrane and with the endoplasmic reticulum. Reduced amounts of the protein were associated with other membranes in the cytoplasm. These data show that, in this system, the Bcl-2 protein associates with the nuclear membrane and intracytoplasmic membranes but is not preferentially associated with mitochondria.
Givol
I
I
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201.
Tsarfaty
I
I
Resau
J
J
Rulong
S
S
da Silva
P P
PP
Nasioulas
G
G
DuHadaway
J
J
Hughes
S H
SH
Ewert
D L
DL
D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry*; D002642:Chick Embryo; D018076:DNA, Complementary / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D005822:Genetic Vectors; D007425:Intracellular Membranes / Q000737:chemistry*; D011518:Proto-Oncogene Proteins / Q000032:analysis* / Q000096:biosynthesis; D019253:Proto-Oncogene Proteins c-bcl-2; D012190:Retroviridae / Q000235:genetics*
D018076:DNA, Complementary; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2