Molecular cloning and regulation of murine fatty acid synthase mRNA

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6074800

Paulauskis, J D; Sul, H S

Mouse liver mRNA that was enriched in sequences coding for fatty acid synthase (FAS) by sucrose-density gradient centrifugation was used as a template for cDNA synthesis. Double-stranded cDNA sequences were inserted into pBR322 and lambdagt10 and cloned. Clones containing putative cDNA sequences for FAS were identified by differential hybridization where /sup 32/P-cDNAs, synthesized from sucrose gradient purified liver mRNA from mice starved or starved and refed a fat-free diet, were used as probes. Two of these clones were further studied and found to contain sequences complementary to FAS mRNA by hybrid-selected translation and specific immunoprecipitation. Using these clones as probes, they selected 33 additional clones containing cDNA sequences for FAS. Partial DNA sequence data for these clones were obtained. Northern blot analysis revealed a single mRNA size of 9.3 kb when a cDNA clone with a 3.1 kb insert was used as a probe. This is in contrast to rat liver FAS which showed two mRNAs sizes of 9.2 and 10.0 kb. They also studied FAS mRNA level of 3T3-L1 preadipocytes during differentiation into adipocytes. An approximate 10-fold increase in FAS mRNA content was observed which corresponded with an increased rate of FAS synthesis indicating pretranslational regulation. The FAS cDNA probe was also employed to demonstrate that induction of FAS in the livers of previously starved mice that were fed a fat-free diet was controlled pretranslationally by a parallel modulation of the FAS mRNA concentration.

• Research Organization: Harvard School of Public Health, Boston, MA
• OSTI ID: 6074800
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOSYNTHESIS
BODY
CARBOXYLIC ACIDS
CELL DIFFERENTIATION
CLONING
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DNA REPLICATION
DNA SEQUENCING
DNA-CLONING
ENZYMES
FASTING
FAT CELLS
GLANDS
HYBRIDIZATION
IMMUNOASSAY
ISOTOPE APPLICATIONS
ISOTOPES
LIGASES
LIGHT NUCLEI
LIVER
MAMMALS
MESSENGER-RNA
MICE
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RNA
RODENTS
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
SYNTHESIS
TRACER TECHNIQUES
VERTEBRATES