In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates.
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 penta...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
Elsevier
2015
|
| _version_ | 1797100467051823104 |
|---|---|
| author | Wawiórka, L Krokowski, D Gordiyenko, Y Krowarsch, D Robinson, C Adam, I Grankowski, N Tchórzewski, M |
| author_facet | Wawiórka, L Krokowski, D Gordiyenko, Y Krowarsch, D Robinson, C Adam, I Grankowski, N Tchórzewski, M |
| author_sort | Wawiórka, L |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T05:37:56Z |
| format | Journal article |
| id | oxford-uuid:e48e3a49-571f-4fad-ae6d-93c2f426db2c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:37:56Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:e48e3a49-571f-4fad-ae6d-93c2f426db2c2022-03-27T10:17:32ZIn vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:e48e3a49-571f-4fad-ae6d-93c2f426db2cEnglishSymplectic Elements at OxfordElsevier2015Wawiórka, LKrokowski, DGordiyenko, YKrowarsch, DRobinson, CAdam, IGrankowski, NTchórzewski, MBACKGROUND: 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. |
| spellingShingle | Wawiórka, L Krokowski, D Gordiyenko, Y Krowarsch, D Robinson, C Adam, I Grankowski, N Tchórzewski, M In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title | In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title_full | In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title_fullStr | In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title_full_unstemmed | In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title_short | In vivo formation of Plasmodium falciparum ribosomal stalk - a unique mode of assembly without stable heterodimeric intermediates. |
| title_sort | in vivo formation of plasmodium falciparum ribosomal stalk a unique mode of assembly without stable heterodimeric intermediates |
| work_keys_str_mv | AT wawiorkal invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT krokowskid invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT gordiyenkoy invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT krowarschd invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT robinsonc invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT adami invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT grankowskin invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates AT tchorzewskim invivoformationofplasmodiumfalciparumribosomalstalkauniquemodeofassemblywithoutstableheterodimericintermediates |