PMID- 6090458
OWN - NLM
STAT- MEDLINE
DCOM- 19841114
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 259
IP  - 19
DP  - 1984 Oct 10
TI  - Identification of a cAMP regulatory region in the gene for rat cytosolic 
      phosphoenolpyruvate carboxykinase (GTP). Use of chimeric genes transfected into 
      hepatoma cells.
PG  - 12161-9
AB  - cAMP stimulates the transcription of the gene for the cytosolic form of 
      phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) in rat liver. We 
      have investigated the nucleotide sequences required for regulation of PEPCK gene 
      expression by cAMP. A chimeric gene was constructed in which a 620-base pair 
      fragment of the 5'-end of the PEPCK gene (including 547 base pairs of 5'-flanking 
      sequence) was ligated to the herpes simplex virus thymidine kinase (TK) 
      structural gene. The PEPCK promoter fragment was introduced either in the proper 
      orientation for transcription of the TK gene or in the opposite orientation. 
      These fusion genes and the parent vector, pOPF, which contains the intact TK 
      gene, were transfected individually into TK-deficient FTO-2B rat hepatoma cells. 
      FTO-2B cells contain an active endogenous PEPCK gene which is stimulated by cAMP. 
      Cells were selected in HAT medium and grown either as mass cell cultures or as 
      individual clones. Dibutyryl cyclic AMP (Bt2cAMP) plus theophylline (16 h) 
      stimulated TK activity 1.6-6.1-fold in cell lines transfected with the PEPCK-TK 
      fusion gene containing the PEPCK promoter fragment in the correct orientation. 
      However, the intact TK gene was not induced by Bt2cAMP after transfection, nor 
      was there any expression of the PEPCK-TK fusion gene in cells which contained the 
      PEPCK promoter fragment in the wrong transcriptional orientation. Bt2cAMP also 
      increased the levels of TK mRNA in cells transfected with the PEPCK-TK fusion 
      gene, but not in cells transfected with the intact TK gene. The chimeric PEPCK-TK 
      mRNA initiated at the PEPCK start site, as determined by S1 nuclease mapping. 
      There was no relationship between the number of copies of the PEPCK-TK gene 
      integrated in the various cell lines and either the basal level of TK activity or 
      its inducibility of Bt2cAMP.
FAU - Wynshaw-Boris, A
AU  - Wynshaw-Boris A
FAU - Lugo, T G
AU  - Lugo TG
FAU - Short, J M
AU  - Short JM
FAU - Fournier, R E
AU  - Fournier RE
FAU - Hanson, R W
AU  - Hanson RW
LA  - eng
SI  - GENBANK/K02299
GR  - AM 24451/AM/NIADDK NIH HHS/United States
GR  - AM07319/AM/NIADDK NIH HHS/United States
GR  - GM 26449/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - E0399OZS9N (Cyclic AMP)
RN  - EC 2.7.1.21 (Thymidine Kinase)
RN  - EC 3.1.21.- (DNA Restriction Enzymes)
RN  - EC 4.1.1.32 (Phosphoenolpyruvate Carboxykinase (GTP))
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - *Chimera
MH  - Cyclic AMP/*physiology
MH  - DNA Restriction Enzymes/metabolism
MH  - *Gene Expression Regulation
MH  - Guanosine Triphosphate
MH  - Liver/enzymology
MH  - Liver Neoplasms, Experimental/*enzymology
MH  - Operon
MH  - Phosphoenolpyruvate Carboxykinase (GTP)/*genetics
MH  - Plasmids
MH  - Rats
MH  - Salmon
MH  - Thymidine Kinase/metabolism
MH  - Transcription, Genetic
MH  - Transfection
EDAT- 1984/10/10 00:00
MHDA- 1984/10/10 00:01
CRDT- 1984/10/10 00:00
PHST- 1984/10/10 00:00 [pubmed]
PHST- 1984/10/10 00:01 [medline]
PHST- 1984/10/10 00:00 [entrez]
AID - S0021-9258(20)71334-7 [pii]
PST - ppublish
SO  - J Biol Chem. 1984 Oct 10;259(19):12161-9.