Selective overproduction of human dihydrofolate reductase in a methotrexate-resistant human-mouse somatic cell hybrid

K. J. Sastry; Teh sheng Chan; Lewis V. Rodriguez

doi:10.1016/0006-291X(85)91202-1

Selective overproduction of human dihydrofolate reductase in a methotrexate-resistant human-mouse somatic cell hybrid

K. J. Sastry, Teh sheng Chan, Lewis V. Rodriguez

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

The development of methotrexate (MTX) resistance in cultured cells results in increased levels of the drug's target enzyme dihydrofolate reductase (DHFR). Stepwise-selected MTX-resistant sublines originating from an MTX-sensitive human-mouse hybrid expressed elevated DHFR levels and human-DHFR specific gene sequence amplification. By high resolution two-dimensional polyacrylamide gradient electrophoresis, human DHFR was shown to be selectively overproduced in VB2a-100 MTX-resistant cells whereas mouse DHFR protein "spots" present in MTX-sensitive parental hybrid were absent in these cells exhibiting 100 μM MTX resistance. These findings and those in a parallel study indicate that concurrent with overproduction of human DHFR and amplification DHFR sequences in VB2a-100, a loss of mouse-specific DHFR gene sequences occurred.

Original language	English (US)
Pages (from-to)	795-803
Number of pages	9
Journal	Biochemical and Biophysical Research Communications
Volume	132
Issue number	2
DOIs	https://doi.org/10.1016/0006-291X(85)91202-1
State	Published - Oct 30 1985
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0006-291X(85)91202-1

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title = "Selective overproduction of human dihydrofolate reductase in a methotrexate-resistant human-mouse somatic cell hybrid",

abstract = "The development of methotrexate (MTX) resistance in cultured cells results in increased levels of the drug's target enzyme dihydrofolate reductase (DHFR). Stepwise-selected MTX-resistant sublines originating from an MTX-sensitive human-mouse hybrid expressed elevated DHFR levels and human-DHFR specific gene sequence amplification. By high resolution two-dimensional polyacrylamide gradient electrophoresis, human DHFR was shown to be selectively overproduced in VB2a-100 MTX-resistant cells whereas mouse DHFR protein {"}spots{"} present in MTX-sensitive parental hybrid were absent in these cells exhibiting 100 μM MTX resistance. These findings and those in a parallel study indicate that concurrent with overproduction of human DHFR and amplification DHFR sequences in VB2a-100, a loss of mouse-specific DHFR gene sequences occurred.",

author = "Sastry, {K. J.} and Chan, {Teh sheng} and Rodriguez, {Lewis V.}",

note = "Funding Information: investigation was supported in part by NIH grants GM 26522 (TSC) and (LVR). The authors wish to acknowledge Rebecca Knowles for technical and Shirlee Mayer for manuscript preparation. Special thanks to S. Ramagli help in photography.",

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doi = "10.1016/0006-291X(85)91202-1",

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T1 - Selective overproduction of human dihydrofolate reductase in a methotrexate-resistant human-mouse somatic cell hybrid

AU - Sastry, K. J.

AU - Chan, Teh sheng

AU - Rodriguez, Lewis V.

N1 - Funding Information: investigation was supported in part by NIH grants GM 26522 (TSC) and (LVR). The authors wish to acknowledge Rebecca Knowles for technical and Shirlee Mayer for manuscript preparation. Special thanks to S. Ramagli help in photography.

PY - 1985/10/30

Y1 - 1985/10/30

N2 - The development of methotrexate (MTX) resistance in cultured cells results in increased levels of the drug's target enzyme dihydrofolate reductase (DHFR). Stepwise-selected MTX-resistant sublines originating from an MTX-sensitive human-mouse hybrid expressed elevated DHFR levels and human-DHFR specific gene sequence amplification. By high resolution two-dimensional polyacrylamide gradient electrophoresis, human DHFR was shown to be selectively overproduced in VB2a-100 MTX-resistant cells whereas mouse DHFR protein "spots" present in MTX-sensitive parental hybrid were absent in these cells exhibiting 100 μM MTX resistance. These findings and those in a parallel study indicate that concurrent with overproduction of human DHFR and amplification DHFR sequences in VB2a-100, a loss of mouse-specific DHFR gene sequences occurred.

AB - The development of methotrexate (MTX) resistance in cultured cells results in increased levels of the drug's target enzyme dihydrofolate reductase (DHFR). Stepwise-selected MTX-resistant sublines originating from an MTX-sensitive human-mouse hybrid expressed elevated DHFR levels and human-DHFR specific gene sequence amplification. By high resolution two-dimensional polyacrylamide gradient electrophoresis, human DHFR was shown to be selectively overproduced in VB2a-100 MTX-resistant cells whereas mouse DHFR protein "spots" present in MTX-sensitive parental hybrid were absent in these cells exhibiting 100 μM MTX resistance. These findings and those in a parallel study indicate that concurrent with overproduction of human DHFR and amplification DHFR sequences in VB2a-100, a loss of mouse-specific DHFR gene sequences occurred.

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Selective overproduction of human dihydrofolate reductase in a methotrexate-resistant human-mouse somatic cell hybrid

Abstract

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