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Evidence for a Functional Change in the Plasma Membrane of Murine Sarcoma Virus-infected Mouse Embryo Cells

TRANSPORT AND TRANSPORT-ASSOCIATED PHOSPHORYLATION OF ¹⁴C-2-DEOXY-d-GLUCOSE

From the ¹ From the Flow Laboratories, Inc., Rockville, Maryland 20852

Enhancement of sugar uptake occurs in murine sarcoma virus-infected mouse cells at the time of transformation. Direct analysis of the sugar transport and transport-associated phosphorylation system of mouse cells infected with the Harvey strain of MSV was performed with 2-deoxy-d-glucose-¹⁴C. It was shown that 2-deoxy-d-glucose-¹⁴C was taken up at various concentrations (10^-2 to 10^-6 m) at a greater rate and was transported with a lower K_m (21.0 to 23.3 x 10^-4 m for uninfected and 3.5 to 5.0 x 10^-4 m for infected cells) and higher V_max (18.2 to 19.2 nmoles per mg of protein per min at 37° for uninfected and 52.3 to 83.5 nmoles per mg of protein per min for infected cells) in the infected cells. The transported substrate was found mainly as a phosphorylated product, 2-deoxy-d-glucose 6-phosphate. The increased phosphorylation of 2-deoxy-d-glucose-¹⁴C by the infected cells was obtained only with intact cells, but not with cell homogenates, indicating that this phosphorylation is not due to increased intracellular hexokinase activity but rather based on a membrane-bound transport system. These data support the hypothesis that alterations in a plasma membrane sugar transport (penetration plus phosphorylation) system occurred during cellular transformation by murine sarcoma virus.

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