Isolation and characterization of the mouse cytosolic phosphoenolpyruvate carboxykinase (GTP) gene: evidence for tissue-specific hypersensitive sites.
A 72 kilobase pair DNA fragment that contains the mouse phosphoenolpyruvate carboxykinase (PEPCK) gene locus, pck1, was isolated from a genomic bacterial artificial chromosome library. The region from approximately -5.5 to +6.6 kilobase pairs relative to the pck1 transcription start site was sequenced and exhibits a high degree of homology to the rat and human genes. Additionally, the chromatin structure of the PEPCK gene in mouse liver resembles that seen in rat. Backcross panel analysis of a microsatellite sequence confirms that the gene is located on chromosome 2. Hypersensitive site analysis was performed on nuclei isolated from the adipocyte cell line 3T3-F442A in the preadipose and adipose states. Several hypersensitive sites are present in the undifferentiated 3T3-F442A cells, before PEPCK mRNA is detected. The same sites are present after differentiation, however, the sensitivity of mHS 3 increases relative to the others. We conclude that the chromatin is open in 3T3-F442A cells and that factors are able to bind in the undifferentiated state but that something else is required for transcription.
Williams
C P
CP
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Postic
C
C
Robin
D
D
Robin
P
P
Parrinello
J
J
Shelton
K
K
Printz
R L
RL
Magnuson
M A
MA
Granner
D K
DK
Forest
C
C
Chalkley
R
R
D016475:3T3 Cells; D017667:Adipocytes / Q000201:enzymology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D015698:Genomic Library; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic
D012333:RNA, Messenger; D011994:Recombinant Proteins; D010729:Phosphoenolpyruvate Carboxykinase (GTP)