Complex environment of nascent polypeptide chains.
Nascent polypeptides enter into high molecular weight complexes with other proteins during chain elongation in vitro and in vivo. The nature of these complexes was investigated using an in vitro translation system programmed with a single mRNA lacking a translational termination codon. Complexes containing nascent polypeptides (molecular mass < 20 kDa), the molecular chaperone hsp 73 and other unidentified proteins can be released from the translationally arrested polysomes by puromycin treatment. The apparent native molecular mass of the nascent chain binding complex was determined to be > 700 kDa by gel-filtration analysis. Complexes between the nascent polypeptide and at least hsp 73 appear to be sensitive to (disrupted by) ATP. The presence of ATP also dramatically alters the sensitivity of the nascent polypeptide chains to digestion by exogenous protease. Collectively, our data indicate that there may be a cytoplasmic machinery, including hsp 70, which comprises a nascent polypeptide chain binding complex.
Hansen
W J
WJ
Department of Medicine, University of California, San Francisco 94143.
Lingappa
V R
VR
Welch
W J
WJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism*; D002918:Chymotrypsin / Q000378:metabolism; D003062:Codon; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D010441:Peptide Chain Elongation, Translational; D010455:Peptides / Q000378:metabolism*; D011132:Polyribosomes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D013728:Terminator Regions, Genetic
D003062:Codon; D018840:HSP70 Heat-Shock Proteins; D010455:Peptides; D012333:RNA, Messenger; D000255:Adenosine Triphosphate; D015500:Chloramphenicol O-Acetyltransferase; D002918:Chymotrypsin