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Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation

BACKGROUND: During Xenopus oocyte maturation, the amount of a 48 kDa protein detected in the 26S proteasome fraction (p48) decreased markedly during oocyte maturation to the low levels seen in unfertilized eggs. The results indicate that the interaction of at least one protein with the 26S proteasom...

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Autores principales: Tokumoto, Toshinobu, Kondo, Ayami, Miwa, Junko, Horiguchi, Ryo, Tokumoto, Mika, Nagahama, Yoshitaka, Okida, Noriyuki, Ishikawa, Katsutoshi
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC179889/
https://www.ncbi.nlm.nih.gov/pubmed/12864926
http://dx.doi.org/10.1186/1471-2091-4-6
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author Tokumoto, Toshinobu
Kondo, Ayami
Miwa, Junko
Horiguchi, Ryo
Tokumoto, Mika
Nagahama, Yoshitaka
Okida, Noriyuki
Ishikawa, Katsutoshi
author_facet Tokumoto, Toshinobu
Kondo, Ayami
Miwa, Junko
Horiguchi, Ryo
Tokumoto, Mika
Nagahama, Yoshitaka
Okida, Noriyuki
Ishikawa, Katsutoshi
author_sort Tokumoto, Toshinobu
collection PubMed
description BACKGROUND: During Xenopus oocyte maturation, the amount of a 48 kDa protein detected in the 26S proteasome fraction (p48) decreased markedly during oocyte maturation to the low levels seen in unfertilized eggs. The results indicate that the interaction of at least one protein with the 26S proteasome changes during oocyte maturation and early development. An alteration in proteasome function may be important for the regulation of developmental events, such as the rapid cell cycle, in the early embryo. In this study, we identified p48. RESULTS: p48 was purified by conventional column chromatography. The resulting purified fraction contained two other proteins with molecular masses of 30 (p30) and 37 (p37) kDa. cDNAs encode elongation factor-1γ and δ were obtained by an immuno-screening method using polyclonal antibodies against purified p48 complex, which recognized p48 and p37. N-terminal amino acid sequence analysis of p30 revealed that it was identical to EF-1β. To identify the p48 complex bound to the 26S proteasome as EF-1βγδ, antibodies were raised against the components of purified p48 complex. Recombinant EF-1 β,γ and δ were expressed in Escherichia coli, and an antibody was raised against purified recombinant EF-1γ. Cross-reactivity of the antibodies toward the p48 complex and recombinant proteins showed it to be specific for each component. These results indicate that the p48 complex bound to the 26S proteasome is the EF-1 complex. MPF phosphorylated EF-1γ was shown to bind to the 26S proteasome. When EF-1γ is phosphorylated by MPF, the association is stabilized. CONCLUSION: p48 bound to the 26S proteasome is identified as the EF-1γ. EF-1 complex is associated with the 26S proteasome in Xenopus oocytes and the interaction is stabilized by MPF-mediated phosphorylation.
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spelling pubmed-1798892003-08-20 Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation Tokumoto, Toshinobu Kondo, Ayami Miwa, Junko Horiguchi, Ryo Tokumoto, Mika Nagahama, Yoshitaka Okida, Noriyuki Ishikawa, Katsutoshi BMC Biochem Research Article BACKGROUND: During Xenopus oocyte maturation, the amount of a 48 kDa protein detected in the 26S proteasome fraction (p48) decreased markedly during oocyte maturation to the low levels seen in unfertilized eggs. The results indicate that the interaction of at least one protein with the 26S proteasome changes during oocyte maturation and early development. An alteration in proteasome function may be important for the regulation of developmental events, such as the rapid cell cycle, in the early embryo. In this study, we identified p48. RESULTS: p48 was purified by conventional column chromatography. The resulting purified fraction contained two other proteins with molecular masses of 30 (p30) and 37 (p37) kDa. cDNAs encode elongation factor-1γ and δ were obtained by an immuno-screening method using polyclonal antibodies against purified p48 complex, which recognized p48 and p37. N-terminal amino acid sequence analysis of p30 revealed that it was identical to EF-1β. To identify the p48 complex bound to the 26S proteasome as EF-1βγδ, antibodies were raised against the components of purified p48 complex. Recombinant EF-1 β,γ and δ were expressed in Escherichia coli, and an antibody was raised against purified recombinant EF-1γ. Cross-reactivity of the antibodies toward the p48 complex and recombinant proteins showed it to be specific for each component. These results indicate that the p48 complex bound to the 26S proteasome is the EF-1 complex. MPF phosphorylated EF-1γ was shown to bind to the 26S proteasome. When EF-1γ is phosphorylated by MPF, the association is stabilized. CONCLUSION: p48 bound to the 26S proteasome is identified as the EF-1γ. EF-1 complex is associated with the 26S proteasome in Xenopus oocytes and the interaction is stabilized by MPF-mediated phosphorylation. BioMed Central 2003-07-16 /pmc/articles/PMC179889/ /pubmed/12864926 http://dx.doi.org/10.1186/1471-2091-4-6 Text en Copyright © 2003 Tokumoto et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Research Article
Tokumoto, Toshinobu
Kondo, Ayami
Miwa, Junko
Horiguchi, Ryo
Tokumoto, Mika
Nagahama, Yoshitaka
Okida, Noriyuki
Ishikawa, Katsutoshi
Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title_full Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title_fullStr Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title_full_unstemmed Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title_short Regulated interaction between polypeptide chain elongation factor-1 complex with the 26S proteasome during Xenopus oocyte maturation
title_sort regulated interaction between polypeptide chain elongation factor-1 complex with the 26s proteasome during xenopus oocyte maturation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC179889/
https://www.ncbi.nlm.nih.gov/pubmed/12864926
http://dx.doi.org/10.1186/1471-2091-4-6
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