Hepatic polysomes that contain apoprotein B mRNA have unusual physical properties.
To characterize the association of the mRNA for apoprotein B (apoB) with ribosomes, rat hepatic cytoplasmic extracts were fractionated by density gradient centrifugation. On linear sucrose gradients, the sedimentation velocity of the 14.4-kilobase apoB mRNA was retarded compared to the mRNAs for other hepatic proteins, which were concentrated in fractions containing the bulk of the polysomes. This unusual distribution of apoB mRNA could not be explained by cotranslational association of nascent apoB peptides with lipids, based on experiments using either detergents to delipidate proteins or puromycin to release nascent peptides from polysomes. The results were also not the result of the editing of apoB mRNA, since the sucrose gradient distributions of both edited and nonedited forms were similar. In contrast, the distribution of a 3'-truncated apoB mRNA (apoB-42, 5.8 kilobases) expressed in rat hepatoma cells resembled that of mRNA of a typical hepatic protein. As opposed to the sedimentation velocity results, on equilibrium density gradients most hepatic apoB mRNA was found in the fraction that contained polysomes. Based on these data, the elongation rate of nascent apoB, and the calculated translational yield of apoB mRNA, we conclude that the majority of rat hepatic apoB mRNA must be part of polysomal complexes with unusual physical properties related to the presence of sequence(s) in the 3'-region of the message. These sequences may either be primary determinants of structural features or binding sites for protein factors that effect conformational changes.
Chen
X
X
Department of Biochemistry, Medical College of Pennsylvania, Philadelphia 19129.
Sparks
J D
JD
Yao
Z
Z
Fisher
E A
EA
D000818:Animals; D001055:Apolipoproteins B / Q000737:chemistry / Q000235:genetics*; D003593:Cytoplasm / Q000737:chemistry; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008099:Liver / Q000737:chemistry* / Q000502:physiology; D008297:Male; D011132:Polyribosomes / Q000737:chemistry* / Q000502:physiology; D017393:RNA Editing; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D013395:Sucrose; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; D008055:Lipids; D012333:RNA, Messenger; D013395:Sucrose