Preparation of functional acetyl-CoA carboxylase mRNA from rat mammary gland.
Poly(A)+ RNA from lactating rat mammary glands was fractionated according to size by isokinetic sucrose gradient centrifugation to obtain a fraction enriched for acetyl-CoA carboxylase. In vitro translation of this RNA preparation yielded apparent full-length acetyl-CoA carboxylase with a molecular weight of 260,000. The synthesized protein was identified as acetyl-CoA carboxylase by specific immunoprecipitation. Tests with antiserum to fatty acid synthetase, revealed that the fractions containing acetyl-CoA carboxylase mRNA also contained mRNA for fatty acid synthetase; both of these mRNAs were approximately 10 kb. Fatty acid synthetase with a molecular weight of 250,000 was synthesized. Using an in vitro rabbit reticulocyte lysate translation system, we have shown that the amount of translatable acetyl-CoA carboxylase mRNA increases during lactation. On the fifth day postpartum the level of translatable acetyl-CoA carboxylase mRNA increased to a peak level seven times that on the day of parturition.
D000103:Acetyl-CoA Carboxylase / Q000235:genetics*; D000818:Animals; D064429:Fatty Acid Synthases / Q000235:genetics; D005260:Female; D007774:Lactation; D008025:Ligases / Q000235:genetics*; D008321:Mammary Glands, Animal / Q000032:analysis*; D008970:Molecular Weight; D011247:Pregnancy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D012333:RNA, Messenger; D064429:Fatty Acid Synthases; D008025:Ligases; D000103:Acetyl-CoA Carboxylase