Regulation of catabolism of microinjected ribonuclease A requires the amino-terminal 20 amino acids

Jonathan M. Backer, L. Bourret, J. F. Dice

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Abstract

RNase A introduced into the cytoplasm of IMR-90 human diploid fibroblasts by erythrocyte-mediated microinjection is degraded with a half-life of ~75 hr in the presence of fetal bovine serum. In response to serum deprivation the degradative rate of microinjected RNase A is enhanced 2-fold. RNase S protein (amino acids 21-124) is degraded with a half-life similar to that of RNase A in the presence of serum, but its catabolism is not increased during serum withdrawal. Reconstitution of RNase S protein with RNase S peptide (amino acids 1-20) restored full enzymatic activity to the S protein as well as the ability of fibroblasts to increase its catabolism during serum deprivation. Finally, RNase S peptide microinjected alone shows the full 2-fold increase in degradative rate during serum withdrawal. These results show that recognition of RNase A for enhanced breakdown during serum deprivation is based on some feature of its amino-terminal 20 amino acids. Furthermore, our results indicate that the enhanced protein catabolism during serum deprivation can be highly selective.

Original languageEnglish (US)
Pages (from-to)2166-2170
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume80
Issue number8 I
StatePublished - 1983
Externally publishedYes

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Pancreatic Ribonuclease
Amino Acids
Serum
Half-Life
Fibroblasts
Proteins
Protein S
Microinjections
Diploidy
Cytoplasm
Erythrocytes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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abstract = "RNase A introduced into the cytoplasm of IMR-90 human diploid fibroblasts by erythrocyte-mediated microinjection is degraded with a half-life of ~75 hr in the presence of fetal bovine serum. In response to serum deprivation the degradative rate of microinjected RNase A is enhanced 2-fold. RNase S protein (amino acids 21-124) is degraded with a half-life similar to that of RNase A in the presence of serum, but its catabolism is not increased during serum withdrawal. Reconstitution of RNase S protein with RNase S peptide (amino acids 1-20) restored full enzymatic activity to the S protein as well as the ability of fibroblasts to increase its catabolism during serum deprivation. Finally, RNase S peptide microinjected alone shows the full 2-fold increase in degradative rate during serum withdrawal. These results show that recognition of RNase A for enhanced breakdown during serum deprivation is based on some feature of its amino-terminal 20 amino acids. Furthermore, our results indicate that the enhanced protein catabolism during serum deprivation can be highly selective.",
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T1 - Regulation of catabolism of microinjected ribonuclease A requires the amino-terminal 20 amino acids

AU - Backer, Jonathan M.

AU - Bourret, L.

AU - Dice, J. F.

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N2 - RNase A introduced into the cytoplasm of IMR-90 human diploid fibroblasts by erythrocyte-mediated microinjection is degraded with a half-life of ~75 hr in the presence of fetal bovine serum. In response to serum deprivation the degradative rate of microinjected RNase A is enhanced 2-fold. RNase S protein (amino acids 21-124) is degraded with a half-life similar to that of RNase A in the presence of serum, but its catabolism is not increased during serum withdrawal. Reconstitution of RNase S protein with RNase S peptide (amino acids 1-20) restored full enzymatic activity to the S protein as well as the ability of fibroblasts to increase its catabolism during serum deprivation. Finally, RNase S peptide microinjected alone shows the full 2-fold increase in degradative rate during serum withdrawal. These results show that recognition of RNase A for enhanced breakdown during serum deprivation is based on some feature of its amino-terminal 20 amino acids. Furthermore, our results indicate that the enhanced protein catabolism during serum deprivation can be highly selective.

AB - RNase A introduced into the cytoplasm of IMR-90 human diploid fibroblasts by erythrocyte-mediated microinjection is degraded with a half-life of ~75 hr in the presence of fetal bovine serum. In response to serum deprivation the degradative rate of microinjected RNase A is enhanced 2-fold. RNase S protein (amino acids 21-124) is degraded with a half-life similar to that of RNase A in the presence of serum, but its catabolism is not increased during serum withdrawal. Reconstitution of RNase S protein with RNase S peptide (amino acids 1-20) restored full enzymatic activity to the S protein as well as the ability of fibroblasts to increase its catabolism during serum deprivation. Finally, RNase S peptide microinjected alone shows the full 2-fold increase in degradative rate during serum withdrawal. These results show that recognition of RNase A for enhanced breakdown during serum deprivation is based on some feature of its amino-terminal 20 amino acids. Furthermore, our results indicate that the enhanced protein catabolism during serum deprivation can be highly selective.

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