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BACKGROUND: The ribosomal stalk composed of P-proteins constitutes a structure on the large ribosomal particle responsible for recruitment of translation factors and stimulation of factor-dependent GTP hydrolysis during translation. The main components of the stalk are P-proteins, which form a pentamer. Despite the conserved basic function of the stalk, the P-proteins do not form a uniform entity, displaying heterogeneity in the primary structure across the eukaryotic lineage. The P-proteins from protozoan parasites are among the most evolutionarily divergent stalk proteins. METHODS: We have assembled P-stalk complex of Plasmodium falciparum in vivo in bacterial system using tricistronic expression cassette and provided its characteristics by biochemical and biophysical methods. RESULTS: All three individual P-proteins, namely uL10/P0, P1 and P2, are indispensable for acquisition of a stable structure of the P stalk complex and the pentameric uL10/P0-(P1-P2)₂form represents the most favorable architecture for parasite P-proteins. CONCLUSION: The formation of P. falciparum P-stalk is driven by trilateral interaction between individual elements which represents unique mode of assembling, without stable P1-P2 heterodimeric intermediate. GENERAL SIGNIFICANCE: On the basis of our mass-spectrometry analysis supported by the bacterial two-hybrid assay and biophysical analyses, a unique pathway of the parasite stalk assembling has been proposed. We suggest that the absence of P1/P2 heterodimer, and the formation of a stable pentamer in the presence of all three proteins, indicate a one-step formation to be the main pathway for the vital ribosomal stalk assembly, whereas the P2 homo-oligomer may represent an off-pathway product with physiologically important nonribosomal role.

Original publication

DOI

10.1016/j.bbagen.2014.10.015

Type

Journal article

Journal

Biochim Biophys Acta

Publication Date

01/2015

Volume

1850

Pages

150 - 158

Keywords

Malaria, Protein complexes, Protein–protein interactions, Ribosomal P proteins, Ribosomal stalk, Ribosomes, Amino Acid Sequence, Circular Dichroism, Electrophoresis, Polyacrylamide Gel, Mass Spectrometry, Models, Molecular, Molecular Sequence Data, Phosphoproteins, Plasmodium falciparum, Protein Binding, Protein Multimerization, Protein Structure, Tertiary, Protozoan Proteins, Recombinant Proteins, Ribosomal Proteins, Ribosomes, Sequence Homology, Amino Acid, Two-Hybrid System Techniques