Functional complementation of yeast ribosomal P0 protein with Plasmodium falciparum P0

Aruna, K.; Chakraborty, Tirtha; Rao, Pavitra N.; Santos, Cruz; Ballesta, Juan P. G.; Sharma, Shobhona

doi:10.1016/j.gene.2005.04.007

Cited by 26 publications

(19 citation statements)

References 25 publications

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“…The inclusion of T. cruzi on this analysis showed that TcP0 protein possesses those two putative P1/P2 interaction regions and its localization is in accordance with SPR and Y2H data (Figure 4). However, some of the residues in these regions are not conserved among species, explaining the lack of complete complementation of yeast P0 deletion strains by heterologous P0 proteins (Rodriguez-Gabriel et al, 2000;Aruna et al, 2005). We then analyzed the number of independent interaction sites present in TcP2b using a set of overlapping peptides (15 amino acid long) that cover the complete sequence of the protein (Supplementary Figure 2A).…”

Section: Tcp1amentioning

confidence: 99%

Interaction map of the Trypanosoma cruzi ribosomal P protein complex (stalk) and the elongation factor 2

Smulski¹,

Longhi²,

Ayub³

et al. 2010

J of Molecular Recognition

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The large subunit of the eukaryotic ribosome possesses a long and protruding stalk formed by the ribosomal P proteins. This structure is involved in the translation step of protein synthesis through interaction with the elongation factor 2 (EF-2). The Trypanosoma cruzi stalk complex is composed of four proteins of about 11 kDa, TcP1α, TcP1β, TcP2α, TcP2β and a fifth TcP0 of about 34 kDa. In a previous work, a yeast two-hybrid (Y2H) protein-protein interaction map of T. cruzi ribosomal P proteins was generated. In order to gain new insight into the assembly of the stalk, a complete interaction map was generated by surface plasmon resonance (SPR) and the kinetics of each interaction was calculated. All previously detected interactions were confirmed and new interacting pairs were found, such as TcP1β-TcP2α and TcP1β-TcP2β. Moreover P2 but not P1 proteins were able to homo-oligomerize. In addition, the region comprising amino acids 210-270 on TcP0 was identified as the region interacting with P1/P2 proteins, using Y2H and SPR. The interaction domains on TcP2β were also mapped by SPR identifying two distinct regions. The assembly order of the pentameric complex was assessed by SPR showing the existence of a hierarchy in the association of the different P proteins forming the stalk. Finally, the TcEF-2 gene was identified, cloned, expressed and refolded. Using SPR analysis we showed that TcEF-2 bound with similar affinity to the four P1/P2 ribosomal P proteins of T. cruzi but with reduced affinity to TcP0.

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Section: Tcp1amentioning

confidence: 99%

Interaction map of the Trypanosoma cruzi ribosomal P protein complex (stalk) and the elongation factor 2

Smulski¹,

Longhi²,

Ayub³

et al. 2010

J of Molecular Recognition

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“…P0 functions as a scaffold for the stalk structure by interacting with 28S rRNA, and the remaining P proteins assemble to form the extended stalk via interactions with P0. P0 is a protein conserved across kingdoms, as shown by the ability of P0 genes from worm, mammals, and protozoa to functionally complement Saccharomyces cerevisiae lacking endogenous P0 (28)(29)(30). P0 is the only essential P protein for both in vitro translational activity of yeast ribosomes and cell survival (31,32).…”

mentioning

confidence: 99%

Ribosomal Protein P0 Promotes Potato Virus A Infection and Functions in Viral Translation Together with VPg and eIF(iso)4E

Hafrén

Eskelin

Mäkinen

2013

J Virol

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We report here that the acidic ribosomal protein P0 is a component of the membrane-associated Potato virus A (PVA) ribonucleoprotein complex. As a constituent of the ribosomal stalk, P0 functions in translation. Although the ribosomal stalk proteins P0, P1, P2, and P3 are all important for PVA infection, P0 appears to have a distinct role from those of the other stalk proteins in infection. Our results indicate that P0 also regulates viral RNA functions as an extraribosomal protein. We reported previously that PVA RNA can be targeted by VPg to a specific gene expression pathway that protects the viral RNA from degradation and facilitates its translation. Here, we show that P0 is essential for this activity of VPg, similar to eIF4E/eIF(iso)4E. We also demonstrate that VPg, P0, and eIF(iso)4E synergistically enhance viral translation. Interestingly, the positive effects of VPg and P0 on viral translation were negatively correlated with the cell-to-cell spread of infection, suggesting that these processes may compete for viral RNA.

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“…cerevisiae P1/P2 proteins efficiently [25,26]. Altogether our data indicate that TcP0 possesses two P1/P2 interaction sites and that P1/P2 proteins can associate in pairs (P1 -P2 /P1 -P2 ) but it was not known whether a hierarchy for P1/P2 association to TcP0 exists.…”

Section: The Ribosomal Stalk: Variable Components and Assemblymentioning

confidence: 74%

Ribosomes from Trypanosomatids: Unique Structural and Functional Properties

Ayub¹,

Lapadula²,

Hoebeke³

et al. 2012

Cell-Free Protein Synthesis

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Additional information is available at the end of the chapter http://dx.doi.org/10.5772/48336 IntroductionTrypanosomatids are a monophyletic group of protozoa that diverged early from the eukaryotic lineage, constituting valuable model organisms for studying variability in different highly conserved processes including protein synthesis. Moreover, several species of trypanosomatids are causing agents of endemic diseases in the third world. There are many evidences suggesting that translation in these organisms shows important differences with that of model organisms such as yeast and mammals. These unique features, which have a great potential relevance for both basic and applied research, will be discussed in this chapter. Structural analysis Cryo-electron microscopy map of Trypanosoma cruzi ribosome: Unique features of the rRNAUsing the cryo-electron microscopy (cryo-EM) technique, a 12Å resolution density map of the T. cruzi 80S ribosome has been constructed [1]. The overall structure of the T. cruzi 80S ribosome exhibits well defined small (40S) and large (60S) subunits (Figure 1). Some of the landmark characteristics of the ribosome structure can be identified in the density map. Compared with the 80S ribosome from yeast, both the small and large ribosomal subunits from T. cruzi are larger, mainly due to the size of the ribosomal RNA molecules. T. cruzi rRNA (18S rRNA: 2,315 nt and 28S rRNA: 4,151 nt) is one-fifth larger than yeast rRNA (18S rRNA: 1,798 nt; 25S rRNA: 3,392 nt) in total number of nucleotides.Although the T. cruzi 80S ribosome possesses conserved ribosomal structures, it exhibits many distinctive structural features in both the small and large subunits. Compared with other eukaryotic ribosomes, the T. cruzi ribosomal 40S subunit appears expanded, due to the addition of a large piece of density adjacent to the platform region ( Figure 1). As can be seen in the secondary structure of the T. cruzi 18S rRNA (Figure 2), this extra density must be attributed to two large expansion segments (ES) in domain II of the 18S rRNA, ES6 and ES7, designated as insertions of helices 21 and 26. These are the two largest ES in the T. cruzi 18S rRNA, involving 504 and 147 nucleotides, respectively.

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Functional complementation of yeast ribosomal P0 protein with Plasmodium falciparum P0

Cited by 26 publications

References 25 publications

Interaction map of the Trypanosoma cruzi ribosomal P protein complex (stalk) and the elongation factor 2

Interaction map of the Trypanosoma cruzi ribosomal P protein complex (stalk) and the elongation factor 2

Ribosomal Protein P0 Promotes Potato Virus A Infection and Functions in Viral Translation Together with VPg and eIF(iso)4E

Ribosomes from Trypanosomatids: Unique Structural and Functional Properties

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