PMID- 2185901
OWN - NLM
STAT- MEDLINE
DCOM- 19900612
LR  - 20190510
IS  - 0143-3334 (Print)
IS  - 0143-3334 (Linking)
VI  - 11
IP  - 5
DP  - 1990 May
TI  - Increased drug resistance following retroviral gene transfer of a chimeric 
      P-enolpyruvate carboxykinase (GTP)-bacterial O6-alkylguanine-DNA alkyltransferase 
      gene into NRK cells.
PG  - 737-43
AB  - Transfection of the Escherichia coli ada gene coding for O6-alkylguanine-DNA 
      alkyltransferase results in expression of ada in mammalian cells and transmission 
      of nitrosourea resistance to cells lacking alkyltransferase activity. We have 
      used a replication-incompetent retrovirus to transfer into mammalian cells a 
      chimeric gene consisting of 548 bp of the promoter-regulatory region of the gene 
      for P-enolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) linked to ada. The 
      PEPCK promoter was used because it is inducible and highly expressed in liver and 
      kidney cells both in vitro and in vivo. After retrovirus infection of the rat 
      kidney cell line, NRK, intact proviral DNA was integrated into the genome of 
      cloned cells. Individual NRK clones produced up to 200 units/mg protein of 
      bacterial alkyltransferase activity compared to 65 units/mg protein of mammalian 
      alkyltransferase in the parent cell line. Transcription of ada mRNA originating 
      from the PEPCK promoter was induced with Bt2cAMP or dexamethasone and the 
      combination caused a 4-fold increase in ada mRNA while total alkyltransferase 
      activity was induced up to 2-fold. NRK clones expressing ada had up to 2.0-fold 
      increased resistance to 1,3-bis(2- chloroethyl)-1- nitrosourea. Thus, retroviral 
      gene transfer of the PEPCKada chimeric gene allows efficient and inducible 
      expression of ada with a resulting increase in alkyltransferase activity and 
      nitrosourea drug resistance. This retrovirus may be used to study the role of 
      alkyltransferase in repair of mutagenic DNA lesions in mammalian cells in vivo.
FAU - Lim, I K
AU  - Lim IK
AD  - Department of Medicine, Pew Center for Molecular Nutrition, Case Western Reserve 
      University School of Medicine, Cleveland, OH.
FAU - Dumenco, L L
AU  - Dumenco LL
FAU - Hatzoglou, M
AU  - Hatzoglou M
FAU - Hanson, R W
AU  - Hanson RW
FAU - Gerson, S L
AU  - Gerson SL
LA  - eng
GR  - CA-45609/CA/NCI NIH HHS/United States
GR  - DK-21889/DK/NIDDK NIH HHS/United States
GR  - ESCA-00134/ES/NIEHS NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Carcinogenesis
JT  - Carcinogenesis
JID - 8008055
RN  - 0 (Hormones)
RN  - 0 (Nitrosourea Compounds)
RN  - EC 2.1.1.- (Methyltransferases)
RN  - EC 2.1.1.63 (O(6)-Methylguanine-DNA Methyltransferase)
RN  - EC 4.1.1.32 (Phosphoenolpyruvate Carboxykinase (GTP))
SB  - IM
MH  - Animals
MH  - Blotting, Northern
MH  - Cell Line
MH  - Cells, Cultured
MH  - Drug Resistance/*genetics
MH  - Enzyme Induction
MH  - Epithelial Cells
MH  - Escherichia coli/genetics
MH  - Fibroblasts/cytology
MH  - Gene Expression Regulation
MH  - Hormones/pharmacology
MH  - Methyltransferases/*genetics/metabolism
MH  - Nitrosourea Compounds/pharmacology
MH  - O(6)-Methylguanine-DNA Methyltransferase
MH  - Phosphoenolpyruvate Carboxykinase (GTP)/*genetics
MH  - Promoter Regions, Genetic
MH  - Retroviridae/*genetics
MH  - Salmon
MH  - Transfection/*genetics
EDAT- 1990/05/01 00:00
MHDA- 1990/05/01 00:01
CRDT- 1990/05/01 00:00
PHST- 1990/05/01 00:00 [pubmed]
PHST- 1990/05/01 00:01 [medline]
PHST- 1990/05/01 00:00 [entrez]
AID - 10.1093/carcin/11.5.737 [doi]
PST - ppublish
SO  - Carcinogenesis. 1990 May;11(5):737-43. doi: 10.1093/carcin/11.5.737.