Isolation and characterization of rat intestinal polyribosomes and RNA during absorption of fat. Increased translation in vitro of apo-AIV.
Polyribosomes were isolated from the intestinal mucosa of fasted and fat-fed rats in the presence of ribonuclease inhibitors. Polyribosomes from fat-fed rats were larger and more efficient in incorporating radioactive aminoacids into proteins than those from fasted rats. Total RNA prepared by guanidine-HCl extraction, from the intestine of fasting and fat-fed rats were translated in vitro in a mRNA-dependent rabbit reticulocyte lysate system in the presence of 35S-methionine. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography of the synthesized peptides showed a relative increase in the radioactivity of some peptides of RNA from fat-fed animals and particularly a two fold increase in preapo-AIV indicating that the intestinal apo-AIV synthesis is under transcriptional regulation by the metabolic processes involved in fat transport, that is, triglyceride-rich lipoprotein production.
Pessah
M
M
Salvat
C
C
Amit
N
N
Infante
R
R
D000818:Animals; D001054:Apolipoproteins A / Q000235:genetics*; D002499:Centrifugation, Density Gradient; D004041:Dietary Fats / Q000378:metabolism*; D005215:Fasting; D005502:Food; D007408:Intestinal Absorption; D007422:Intestines / Q000648:ultrastructure*; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D011132:Polyribosomes / Q000032:analysis*; D014176:Protein Biosynthesis; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains
D001054:Apolipoproteins A; D004041:Dietary Fats; C022205:apolipoprotein A-IV; D012313:RNA; D007930:Leucine