PMID:10681589

Citation	Subramanian, A and Miller, DM (2000) Structural analysis of alpha-enolase. Mapping the functional domains involved in down-regulation of the c-myc protooncogene. J. Biol. Chem. 275:5958-65
Abstract	Myc-binding protein-1 (MBP-1) is a 37-kDa protein with sequence homology to the 3' portion of the alpha-enolase gene. alpha-Enolase is a 48-kDa protein, which plays a critical role in the glycolytic pathway. MBP-1 binds to the c-myc P2 promoter and down-regulates c-myc expression. We have investigated the role of alpha-enolase in regulation of the c-myc protooncogene. RNase protection assay shows that alpha-enolase is transcribed into a single RNA species in HeLa cells. A start codon, 400 base pairs downstream of the alpha-enolase ATG, corresponds to the MBP-1 ATG, suggesting that MBP-1 is an alternative translation initiation product of the alpha-enolase RNA. Domain mapping was performed using constructs containing truncations of the alpha-enolase gene. In vitro binding to the c-myc gene was abolished after deletion of the N-terminal portion of alpha-enolase. In order to determine the relationship between DNA binding activity and transcription inhibition, we performed co-transfection assays in HeLa cells. These studies confirmed that an N-terminal deletion of alpha-enolase is unable to down-regulate c-myc promoter activity. Our data suggest that alpha-enolase plays an important role in regulation of c-myc promoter activity in the form of an alternative translation product MBP-1, which is distinct from its role as a glycolytic enzyme.
Links	PubMed
Keywords	Blotting, Northern; DNA-Binding Proteins; Down-Regulation; Gene Deletion; Genes, myc/genetics; HeLa Cells; Humans; Luciferases/metabolism; Mutagenesis, Site-Directed; Neoplasm Proteins/genetics; Neoplasm Proteins/metabolism; Phosphopyruvate Hydratase/chemistry; Phosphopyruvate Hydratase/physiology; Plasmids; Protein Biosynthesis; Protein Isoforms; Protein Structure, Tertiary; RNA, Messenger/metabolism; Ribonucleases/metabolism; Structure-Activity Relationship; Transcription, Genetic; Transfection; Tumor Markers, Biological; Tumor Suppressor Proteins
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:ENOA	involved_in	GO:0045892: negative regulation of transcription, DNA-templated	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:ENOA		GO:0045892 : negative regulation of transcription, DNA-dependent	ECO:0000315:		P		Figure 4, 6, and 8.	complete CACAO 5688

References

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