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Biochim Biophys Acta. 2015 Jan;1850(1):150-8. doi: 10.1016/j.bbagen.2014.10.015. Epub 2014 Oct 23.

In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates.

Author information

1
Department of Molecular Biology, Maria Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland.
2
Department of Chemistry, University of Oxford, South Parks Rd, Oxford OX1 3QZ, UK.
3
Department of Protein Biotechnology, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw, Poland.
4
Department of Obstetrics & Gynecology, Faculty of Medicine, AlKaser Street, University of Khartoum, Khartoum, Sudan.
5
Department of Molecular Biology, Maria Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland. Electronic address: maro@hektor.umcs.lublin.pl.

Abstract

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.

KEYWORDS:

Malaria; Protein complexes; Protein–protein interactions; Ribosomal P proteins; Ribosomal stalk; Ribosomes

PMID:
25450178
DOI:
10.1016/j.bbagen.2014.10.015
[Indexed for MEDLINE]

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