Use of a cloned multidrug resistance gene for coamplification and overproduction of major excreted protein, a transformation-regulated secreted acid protease

Journal Article · Mon Aug 01 04:00:00 EDT 1988 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.8.8.3316· OSTI ID:6737158

Kane, S E; Troen, B R; Gal, S; Ueda, K; Pastan, I; Gottesman, M M

Malignantly transformed mouse fibroblasts synthesize and secrete large amounts of major excreted protein (MEP), a 39,000-dalton precursor to an acid protease (cathepsin L). To evaluate the possible role of this protease in the transformed phenotype, the authors transfected cloned genes for mouse or human MEP into mouse MIH 3T3 cells with an expression vector for the dominant, selectable human multidrug resistance (MDR1) gene. The cotransfected MEP sequences were efficiently coamplified and transcribed during stepwise selection for multidrug resistance in colchicine. The transfected NIH 3T3 cell lines containing amplified MEP sequences synthesized as much MEP as did Kirsten sarcoma virus-transformed NIH 3T3 cells. The MEP synthesized by cells transfected with the cloned mouse and human MEP genes were also secreted. Elevated synthesis and secretion of MEP by NIH 3T3 cells did not change the nontransformed phenotype of these cells.

• Research Organization: Lab. of Molecular Biology, National Cancer Institute, Bethesda, MD (US)
• OSTI ID: 6737158
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 8:8; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ACID PROTEINASES
ANIMAL CELLS
ANIMALS
CELL CULTURES
CELL TRANSFORMATIONS
CLONING
CONNECTIVE TISSUE CELLS
DNA HYBRIDIZATION
DNA-CLONING
DRUGS
ENZYMES
FIBROBLASTS
GENE REGULATION
HYBRIDIZATION
HYDROLASES
MAMMALS
MAN
MICE
MICROORGANISMS
MOLECULAR BIOLOGY
ONCOGENIC TRANSFORMATIONS
ONCOGENIC VIRUSES
PARASITES
PEPTIDE HYDROLASES
PHENOTYPE
PRIMATES
RODENTS
SOMATIC CELLS
VERTEBRATES
VIRUSES