The ribosomal stalk is required for ribosome binding, depurination of the rRNA and cytotoxicity of ricin a chain in Saccharomyces cerevisiae

Jia-chi Chiou, Xiao Ping Li, Miguel Remacha, Juan P.G. Ballesta, Nilgun E. Tumer

Research output: Journal article publicationJournal articleAcademic researchpeer-review

62 Citations (Scopus)


Ribosome inactivating proteins (RIPs) like ricin, pokeweed antiviral protein (PAP) and Shiga-like toxins 1 and 2 (Stx1 and Stx2) share the same substrate, the α-sarcin/ricin loop, but differ in their specificities towards prokaryotic and eukaryotic ribosomes. Ricin depurinates the eukaryotic ribosomes more efficiently than the prokaryotic ribosomes, while PAP can depurinate both types of ribosomes. Accumulating evidence suggests that different docking sites on the ribosome might be used by different RIPs, providing a basis for understanding the mechanism underlying their kingdom specificity. Our previous results demonstrated that PAP binds to the ribosomal protein L3 to depurinate the α-sarcin/ricin loop and binding of PAP to L3 was critical for its cytotoxicity. Here, we used surface plasmon resonance to demonstrate that ricin toxin A chain (RTA) binds to the P1 and P2 proteins of the ribosomal stalk in Saccharomyces cerevisiae. Ribosomes from the P protein mutants were depurinated less than the wild-type ribosomes when treated with RTA in vitro. Ribosome depurination was reduced when RTA was expressed in the ΔP1 and ΔP2 mutants in vivo and these mutants were more resistant to the cytotoxicity of RTA than the wild-type cells. We further show that while RTA, Stx1 and Stx2 have similar requirements for ribosome depurination, PAP has different requirements, providing evidence that the interaction of RIPs with different ribosomal proteins is responsible for their ribosome specificity.
Original languageEnglish
Pages (from-to)1441-1452
Number of pages12
JournalMolecular Microbiology
Issue number6
Publication statusPublished - 1 Dec 2008
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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