Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Débora N. Santos; Pedro H.N. Aguiar; Francisco P. Lobo; Marina M. Mourão; José H.M. Tambor; Analina F. Valadão; Adlane Vilas-Boas; Francisco G. Nobrega; Philip T. LoVerde; Andréa M. Macedo; Sérgio D.J. Pena; Carlos R. Machado; Glória R. Franco

doi:10.1016/j.exppara.2007.02.012

Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Débora N. Santos, Pedro H.N. Aguiar, Francisco P. Lobo, Marina M. Mourão, José H.M. Tambor, Analina F. Valadão, Adlane Vilas-Boas, Francisco G. Nobrega, Philip T. LoVerde, Andréa M. Macedo, Sérgio D.J. Pena, Carlos R. Machado, Glória R. Franco

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1^- yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxΔ24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation.

Original language	English (US)
Pages (from-to)	440-449
Number of pages	10
Journal	Experimental Parasitology
Volume	116
Issue number	4
DOIs	https://doi.org/10.1016/j.exppara.2007.02.012
State	Published - Aug 2007
Externally published	Yes

Keywords

HRT1
RING box
SCF complex
Schistosoma mansoni
SmRbx
Trematode
Ubiquitination

ASJC Scopus subject areas

Infectious Diseases
Parasitology
Immunology

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10.1016/j.exppara.2007.02.012

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Santos, D. N., Aguiar, P. H. N., Lobo, F. P., Mourão, M. M., Tambor, J. H. M., Valadão, A. F., Vilas-Boas, A., Nobrega, F. G., LoVerde, P. T., Macedo, A. M., Pena, S. D. J., Machado, C. R., & Franco, G. R. (2007). Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination. Experimental Parasitology, 116(4), 440-449. https://doi.org/10.1016/j.exppara.2007.02.012

Santos, DN, Aguiar, PHN, Lobo, FP, Mourão, MM, Tambor, JHM, Valadão, AF, Vilas-Boas, A, Nobrega, FG, LoVerde, PT, Macedo, AM, Pena, SDJ, Machado, CR & Franco, GR 2007, 'Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination', Experimental Parasitology, vol. 116, no. 4, pp. 440-449. https://doi.org/10.1016/j.exppara.2007.02.012

@article{2ee381a886004c0396d975edd548e084,

title = "Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination",

abstract = "The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1- yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxΔ24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation.",

keywords = "HRT1, RING box, SCF complex, Schistosoma mansoni, SmRbx, Trematode, Ubiquitination",

author = "Santos, {D{\'e}bora N.} and Aguiar, {Pedro H.N.} and Lobo, {Francisco P.} and Mour{\~a}o, {Marina M.} and Tambor, {Jos{\'e} H.M.} and Valad{\~a}o, {Analina F.} and Adlane Vilas-Boas and Nobrega, {Francisco G.} and LoVerde, {Philip T.} and Macedo, {Andr{\'e}a M.} and Pena, {S{\'e}rgio D.J.} and Machado, {Carlos R.} and Franco, {Gl{\'o}ria R.}",

note = "Funding Information: The authors thank K{\'a}tia Barroso for carrying out automated DNA sequencing, Neuza Antunes Rodrigues for technical support and Dr Carlos Rosa for helpful insights on yeast cell biology. This work received financial support from the following sources: CAPES (Ph.D. scholarship for DNS) and a CNPq grant to GRF (Proc. No. 470425/2001-6).",

year = "2007",

month = aug,

doi = "10.1016/j.exppara.2007.02.012",

language = "English (US)",

volume = "116",

pages = "440--449",

journal = "Experimental Parasitology",

issn = "0014-4894",

publisher = "Academic Press Inc.",

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TY - JOUR

T1 - Schistosoma mansoni

T2 - Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

AU - Santos, Débora N.

AU - Aguiar, Pedro H.N.

AU - Lobo, Francisco P.

AU - Mourão, Marina M.

AU - Tambor, José H.M.

AU - Valadão, Analina F.

AU - Vilas-Boas, Adlane

AU - Nobrega, Francisco G.

AU - LoVerde, Philip T.

AU - Macedo, Andréa M.

AU - Pena, Sérgio D.J.

AU - Machado, Carlos R.

AU - Franco, Glória R.

N1 - Funding Information: The authors thank Kátia Barroso for carrying out automated DNA sequencing, Neuza Antunes Rodrigues for technical support and Dr Carlos Rosa for helpful insights on yeast cell biology. This work received financial support from the following sources: CAPES (Ph.D. scholarship for DNS) and a CNPq grant to GRF (Proc. No. 470425/2001-6).

PY - 2007/8

Y1 - 2007/8

N2 - The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1- yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxΔ24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation.

AB - The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1- yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxΔ24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation.

KW - HRT1

KW - RING box

KW - SCF complex

KW - Schistosoma mansoni

KW - SmRbx

KW - Trematode

KW - Ubiquitination

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U2 - 10.1016/j.exppara.2007.02.012

DO - 10.1016/j.exppara.2007.02.012

M3 - Article

C2 - 17420016

AN - SCOPUS:34249852188

SN - 0014-4894

VL - 116

SP - 440

EP - 449

JO - Experimental Parasitology

JF - Experimental Parasitology

IS - 4

ER -

Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Abstract

Keywords

ASJC Scopus subject areas

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