Induction of human high K(M) 5'-nucleotidase in cultured 293 cells.

Abstract:

:Human 293 cells were stably transfected with a plasmid introducing a receptor for the ecdysone analog muristerone. The cells were further stably transfected with muristerone-inducible expression vectors carrying either the cDNA for the human high K(M) 5'-nucleotidase or the coding sequence of the nucleotidase linked to the 5'-end of the sequence for the green fluorescent protein. Upon induction, both types of transfectants overproduced nucleotidase activity in a time- and dose-dependent manner. Western blots gave values close to the expected subunit molecular masses of 65 and 92 kDa, respectively, excluding processing of the induced proteins. Cells induced to overexpress the nucleotidase showed a decreased growth rate and contained smaller pools of each of the four common ribonucleoside triphosphates. They showed no increased resistance to the toxicity of 2-chlorodeoxyadenosine.

journal_name

Exp Cell Res

authors

Gazziola C,Moras M,Ferraro P,Gallinaro L,Verin R,Rampazzo C,Reichard P,Bianchi V

doi

10.1006/excr.1999.4681

keywords:

subject

Has Abstract

pub_date

1999-12-15 00:00:00

pages

474-82

issue

2

eissn

0014-4827

issn

1090-2422

pii

S0014-4827(99)94681-5

journal_volume

253

pub_type

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