Structural change of ribosomes during apoptosis: degradation and externalization of ribosomal proteins in doxorubicin-treated Jurkat cells.
Changes in the amount and localization of human ribosomal proteins during apoptosis were determined. When total lysates of Jurkat cells undergoing apoptosis induced by doxorubicin were analyzed by Western blotting, degradation of three ribosomal proteins, S18, L5, and L14, was detected at 48 h after the induction of apoptosis. Decreases in the amounts of these three ribosomal proteins were also observed in ribosome-enriched fractions. These changes were partly abolished by the addition of the pan-caspase inhibitor z-VAD-fmk. Moreover, formation of the 80S ribosome complex appeared to be inhibited at 48 h after apoptosis induction. On the other hand, the rate of protein synthesis, assessed by measuring the incorporation of [35S]Met into bulk proteins, decreased as early as 12 h after the addition of doxorubicin. These results indicate that changes in the amount of ribosomal proteins and the overall structure of ribosomes in apoptosing cells occur after protein synthesis declines. Finally, analyses by flow cytometry, immunofluorescence, and Western blotting showed that six ribosomal proteins, S15, P0, L5, L6, L36a, and L41, were relocalized and expressed at the cell surface during apoptosis. The above results collectively indicate that ribosomes are structurally altered in apoptotic cells following inactivation of protein synthesis.
Nishida
Jun
J
Graduate School of Natural Science and Technology, Kanazawa University, Kanazawa, Ishikawa 920-0934, Japan.
Shiratsuchi
Akiko
A
Nadano
Daita
D
Sato
Taka-aki
TA
Nakanishi
Yoshinobu
Y
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D004317:Doxorubicin / Q000494:pharmacology*; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells / Q000187:drug effects*; D021381:Protein Transport / Q000502:physiology; D012269:Ribosomal Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D012270:Ribosomes / Q000502:physiology / Q000648:ultrastructure*
D000970:Antineoplastic Agents; D012269:Ribosomal Proteins; D004317:Doxorubicin