The muscle creatine kinase gene is regulated by multiple upstream elements, including a muscle-specific enhancer
Muscle creatine kinase (MCK) is induced to high levels during skeletal muscle differentiation. The authors examined the upstream regulatory elements of the mouse MCK gene which specify its activation during myogenesis in culture. Fusion genes containing up to 3,300 nucleotides (nt) of MCK 5' flanking DNA in various positions and orientations relative to the bacterial chloramphenicol acetyltransferase (CAT) structural gene were transfected into cultured cells. Transient expression of CAT was compared between proliferating and differentiated MM14 mouse myoblasts and with nonmyogenic mouse L cells. The major effector of high-level expression was found to have the properties of a transcriptional enhancer. This element, located between 1,050 and 1,256 nt upstream of the transcription start site, was also found to have a major influence on the tissue and differentiation specificity of MCK expression; it activated either the MCK promoter or heterologous promoters only in differentiated muscle cells. Comparisons of viral and cellular enhancer sequences with the MCK enhancer revealed some similarities to essential regions of the simian virus 40 enhancer as well as to a region of the immunoglobulin heavy-chain enhancer, which has been implicated in tissue-specific protein binding. Even in the absence of the enhancer, low-level expression from a 776-nt MCK promoter retained differentiation specificity. In addition to positive regulatory elements, our data provide some evidence for negative regulatory elements with activity in myoblasts. These may contribute to the cell type and differentiation specificity of MCK expression.
- Research Organization:
- Dept. of Biochemistry, Univ. of California Medical Center, San Francisco, CA (US)
- OSTI ID:
- 6772518
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 8:1
- Country of Publication:
- United States
- Language:
- English
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CELL DIFFERENTIATION
MOLECULAR BIOLOGY
GENE OPERONS
MUSCLES
GENE REGULATION
PHOSPHOTRANSFERASES
GENES
ACTIVATION ANALYSIS
CELL CULTURES
CREATINE
DNA SEQUENCING
ENZYME ACTIVITY
ENZYME INDUCTION
IMMUNOGLOBULINS
STEM CELLS
AMINO ACIDS
ANIMAL CELLS
CARBOXYLIC ACIDS
CHEMICAL ANALYSIS
ENZYMES
GLOBULINS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES
550200* - Biochemistry
550800 - Morphology