Cis-regulation of the L-type pyruvate kinase gene promoter by glucose, insulin and cyclic AMP

A DNA fragment spanning nucleotides -183 to -4 with respect to the cap site of the rat L-type pyruvate kinase (L-PK) gene contains at least four binding sites for putative transcriptional factors: hepatocyte nuclear factor 1 (HNF1), liver factor A1 (LF-A1), nuclear factor 1 (NF1), and major late transcription factor (MLTF). This fragment was used to direct transcription of a reporter sequence (a G-free cassette) in cell extracts. This L-PK promoter was active in liver nuclear extracts, but not in extracts from nonhepatic tissues. A reduction of 50% of the activity was obtained with a deleted L-PK promoter containing only the HNF1-binding site. In contrast, deletion of the HNF1-binding site inactivated the promoter by more than 90%. These results were confirmed by titration experiments with synthetic oligonucleotides. Titration of HNF1 resulted in an 85% decrease of transcriptional activity, while titration of LF-A1 resulted in only a 40% decrease. The influence of NF1 and MLTF seemed to be marginal in this system. The proximal 5'-flanking sequence of the L-PK gene therefore appears to function in vitro as an efficient liver-specific promoter which requires the binding of the liver factor HNF1 and which is also stimulated by the binding of another liver-specific factor, LF-A1.

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Negative cyclic AMP response elements in the promoter of the L-type pyruvate kinase gene

FEBS Letters ◽

10.1016/s0014-5793(99)01203-x ◽

1999 ◽

Vol 459 (1) ◽

pp. 9-14 ◽

Cited By ~ 14

Author(s):

Laurence Gourdon ◽

Dan Qing Lou ◽

Michel Raymondjean ◽

Mireille Vasseur-Cognet ◽

Axel Kahn

Keyword(s):

Cyclic Amp ◽

Pyruvate Kinase ◽

Kinase Gene ◽

Response Elements

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In vitroandin vivoprotein - DNA interactions on the rat erythroid-specific L′ pyruvate kinase gene promoter

Nucleic Acids Research ◽

10.1093/nar/20.21.5669 ◽

1992 ◽

Vol 20 (21) ◽

pp. 5669-5676 ◽

Cited By ~ 11

Author(s):

Virginie Lacronique ◽

Didier Boquet ◽

Soledad Lopez ◽

Axel Kahn ◽

Michel Raymondjean

Keyword(s):

Pyruvate Kinase ◽

Gene Promoter ◽

Dna Interactions ◽

Kinase Gene

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Analysis by cell-free transcription of the liver-specific pyruvate kinase gene promoter

Molecular and Cellular Biology ◽

10.1128/mcb.9.10.4409-4415.1989 ◽

1989 ◽

Vol 9 (10) ◽

pp. 4409-4415

Author(s):

S Vaulont ◽

N Puzenat ◽

A Kahn ◽

M Raymondjean

Keyword(s):

Binding Site ◽

Pyruvate Kinase ◽

Gene Promoter ◽

Specific Factor ◽

Nuclear Factor ◽

Flanking Sequence ◽

Kinase Gene ◽

Cell Extracts ◽

Nuclear Factor 1

A DNA fragment spanning nucleotides -183 to -4 with respect to the cap site of the rat L-type pyruvate kinase (L-PK) gene contains at least four binding sites for putative transcriptional factors: hepatocyte nuclear factor 1 (HNF1), liver factor A1 (LF-A1), nuclear factor 1 (NF1), and major late transcription factor (MLTF). This fragment was used to direct transcription of a reporter sequence (a G-free cassette) in cell extracts. This L-PK promoter was active in liver nuclear extracts, but not in extracts from nonhepatic tissues. A reduction of 50% of the activity was obtained with a deleted L-PK promoter containing only the HNF1-binding site. In contrast, deletion of the HNF1-binding site inactivated the promoter by more than 90%. These results were confirmed by titration experiments with synthetic oligonucleotides. Titration of HNF1 resulted in an 85% decrease of transcriptional activity, while titration of LF-A1 resulted in only a 40% decrease. The influence of NF1 and MLTF seemed to be marginal in this system. The proximal 5'-flanking sequence of the L-PK gene therefore appears to function in vitro as an efficient liver-specific promoter which requires the binding of the liver factor HNF1 and which is also stimulated by the binding of another liver-specific factor, LF-A1.

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