Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea

Patrícia Diogo De Melo Godoy, Luis Antonio Nogueira-Junior, Lisvane S. Paes, Alberto Cornejo, Rafael Miyazawa Martins, Ariel M. Silber, Sergio Schenkman, M. Carolina Elias

Resultado de la investigación: Article

46 Citas (Scopus)

Resumen

In unicellular eukaryotes, such as Saccharomyces cerevisiae, and in multicellular organisms, the replication origin is recognized by the heterohexamer origin recognition complex (ORC) containing six proteins, Orel to Orc6, while in members of the domain Archaea, the replication origin is recognized by just one protein, Orc1/Cdc6; the sequence of Orc1/Cdc6 is highly related to those of Orel and Cdc6. Similar to Archaea, trypanosomatid genomes contain only one gene encoding a protein named Orel. Since trypanosome Ore1 is also homologous to Cdc6, in this study we named the Ore1 protein from trypanosomes Orc1/Cdc6. Here we show that the recombinant Orc1/Cdc6 from Trypanosoma cruzi (TbOrcl/Cdc6) and from Trypanosoma brucei (TbOrcl/Cdc6) present ATPase activity, typical of prereplication machinery components. Also, TcOrc1/Cdc6 and TbOrc1/Cdc6 replaced yeast Cdc6 but not Orel in a phenotypic complementation assay. The induction of Orc1/Cdc6 silencing by RNA interference in T. brucei resulted in enucleated cells, strongly suggesting the involvement of Orc1/Cdc6 in DNA replication. Orc1/Cdc6 is expressed during the entire cell cycle in the nuclei of trypanosomes, remaining associated with chromatin in all stages of the cell cycle. These results allowed us to conclude that Orcl/Cdc6 is indeed a member of the trypanosome prereplication machinery and point out that trypanosomes carry a prereplication machinery that is less complex than other eukaryotes and closer to archaea.

Idioma originalEnglish
Páginas (desde-hasta)1592-1603
Número de páginas12
PublicaciónEukaryotic Cell
Volumen8
N.º10
DOI
EstadoPublished - oct 2009

Huella dactilar

Trypanosomiasis
Archaea
Trypanosoma brucei brucei
Replication Origin
Proteins
Eukaryota
Cell Cycle
Origin Recognition Complex
Trypanosoma cruzi
RNA Interference
DNA Replication
Chromatin
Saccharomyces cerevisiae
Adenosine Triphosphatases
Yeasts
Genome
Genes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Medicine(all)

Citar esto

De Melo Godoy, P. D., Nogueira-Junior, L. A., Paes, L. S., Cornejo, A., Martins, R. M., Silber, A. M., ... Elias, M. C. (2009). Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea. Eukaryotic Cell, 8(10), 1592-1603. https://doi.org/10.1128/EC.00161-09
De Melo Godoy, Patrícia Diogo ; Nogueira-Junior, Luis Antonio ; Paes, Lisvane S. ; Cornejo, Alberto ; Martins, Rafael Miyazawa ; Silber, Ariel M. ; Schenkman, Sergio ; Elias, M. Carolina. / Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea. En: Eukaryotic Cell. 2009 ; Vol. 8, N.º 10. pp. 1592-1603.
@article{1f67dddc2fee4d46adc9441ccd8d39a2,
title = "Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea",
abstract = "In unicellular eukaryotes, such as Saccharomyces cerevisiae, and in multicellular organisms, the replication origin is recognized by the heterohexamer origin recognition complex (ORC) containing six proteins, Orel to Orc6, while in members of the domain Archaea, the replication origin is recognized by just one protein, Orc1/Cdc6; the sequence of Orc1/Cdc6 is highly related to those of Orel and Cdc6. Similar to Archaea, trypanosomatid genomes contain only one gene encoding a protein named Orel. Since trypanosome Ore1 is also homologous to Cdc6, in this study we named the Ore1 protein from trypanosomes Orc1/Cdc6. Here we show that the recombinant Orc1/Cdc6 from Trypanosoma cruzi (TbOrcl/Cdc6) and from Trypanosoma brucei (TbOrcl/Cdc6) present ATPase activity, typical of prereplication machinery components. Also, TcOrc1/Cdc6 and TbOrc1/Cdc6 replaced yeast Cdc6 but not Orel in a phenotypic complementation assay. The induction of Orc1/Cdc6 silencing by RNA interference in T. brucei resulted in enucleated cells, strongly suggesting the involvement of Orc1/Cdc6 in DNA replication. Orc1/Cdc6 is expressed during the entire cell cycle in the nuclei of trypanosomes, remaining associated with chromatin in all stages of the cell cycle. These results allowed us to conclude that Orcl/Cdc6 is indeed a member of the trypanosome prereplication machinery and point out that trypanosomes carry a prereplication machinery that is less complex than other eukaryotes and closer to archaea.",
author = "{De Melo Godoy}, {Patr{\'i}cia Diogo} and Nogueira-Junior, {Luis Antonio} and Paes, {Lisvane S.} and Alberto Cornejo and Martins, {Rafael Miyazawa} and Silber, {Ariel M.} and Sergio Schenkman and Elias, {M. Carolina}",
year = "2009",
month = "10",
doi = "10.1128/EC.00161-09",
language = "English",
volume = "8",
pages = "1592--1603",
journal = "Eukaryotic Cell",
issn = "1535-9778",
publisher = "American Society for Microbiology",
number = "10",

}

De Melo Godoy, PD, Nogueira-Junior, LA, Paes, LS, Cornejo, A, Martins, RM, Silber, AM, Schenkman, S & Elias, MC 2009, 'Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea', Eukaryotic Cell, vol. 8, n.º 10, pp. 1592-1603. https://doi.org/10.1128/EC.00161-09

Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea. / De Melo Godoy, Patrícia Diogo; Nogueira-Junior, Luis Antonio; Paes, Lisvane S.; Cornejo, Alberto; Martins, Rafael Miyazawa; Silber, Ariel M.; Schenkman, Sergio; Elias, M. Carolina.

En: Eukaryotic Cell, Vol. 8, N.º 10, 10.2009, p. 1592-1603.

Resultado de la investigación: Article

TY - JOUR

T1 - Trypanosome prereplication machinery contains a single functional Orc1/Cdc6 protein, which is typical of archaea

AU - De Melo Godoy, Patrícia Diogo

AU - Nogueira-Junior, Luis Antonio

AU - Paes, Lisvane S.

AU - Cornejo, Alberto

AU - Martins, Rafael Miyazawa

AU - Silber, Ariel M.

AU - Schenkman, Sergio

AU - Elias, M. Carolina

PY - 2009/10

Y1 - 2009/10

N2 - In unicellular eukaryotes, such as Saccharomyces cerevisiae, and in multicellular organisms, the replication origin is recognized by the heterohexamer origin recognition complex (ORC) containing six proteins, Orel to Orc6, while in members of the domain Archaea, the replication origin is recognized by just one protein, Orc1/Cdc6; the sequence of Orc1/Cdc6 is highly related to those of Orel and Cdc6. Similar to Archaea, trypanosomatid genomes contain only one gene encoding a protein named Orel. Since trypanosome Ore1 is also homologous to Cdc6, in this study we named the Ore1 protein from trypanosomes Orc1/Cdc6. Here we show that the recombinant Orc1/Cdc6 from Trypanosoma cruzi (TbOrcl/Cdc6) and from Trypanosoma brucei (TbOrcl/Cdc6) present ATPase activity, typical of prereplication machinery components. Also, TcOrc1/Cdc6 and TbOrc1/Cdc6 replaced yeast Cdc6 but not Orel in a phenotypic complementation assay. The induction of Orc1/Cdc6 silencing by RNA interference in T. brucei resulted in enucleated cells, strongly suggesting the involvement of Orc1/Cdc6 in DNA replication. Orc1/Cdc6 is expressed during the entire cell cycle in the nuclei of trypanosomes, remaining associated with chromatin in all stages of the cell cycle. These results allowed us to conclude that Orcl/Cdc6 is indeed a member of the trypanosome prereplication machinery and point out that trypanosomes carry a prereplication machinery that is less complex than other eukaryotes and closer to archaea.

AB - In unicellular eukaryotes, such as Saccharomyces cerevisiae, and in multicellular organisms, the replication origin is recognized by the heterohexamer origin recognition complex (ORC) containing six proteins, Orel to Orc6, while in members of the domain Archaea, the replication origin is recognized by just one protein, Orc1/Cdc6; the sequence of Orc1/Cdc6 is highly related to those of Orel and Cdc6. Similar to Archaea, trypanosomatid genomes contain only one gene encoding a protein named Orel. Since trypanosome Ore1 is also homologous to Cdc6, in this study we named the Ore1 protein from trypanosomes Orc1/Cdc6. Here we show that the recombinant Orc1/Cdc6 from Trypanosoma cruzi (TbOrcl/Cdc6) and from Trypanosoma brucei (TbOrcl/Cdc6) present ATPase activity, typical of prereplication machinery components. Also, TcOrc1/Cdc6 and TbOrc1/Cdc6 replaced yeast Cdc6 but not Orel in a phenotypic complementation assay. The induction of Orc1/Cdc6 silencing by RNA interference in T. brucei resulted in enucleated cells, strongly suggesting the involvement of Orc1/Cdc6 in DNA replication. Orc1/Cdc6 is expressed during the entire cell cycle in the nuclei of trypanosomes, remaining associated with chromatin in all stages of the cell cycle. These results allowed us to conclude that Orcl/Cdc6 is indeed a member of the trypanosome prereplication machinery and point out that trypanosomes carry a prereplication machinery that is less complex than other eukaryotes and closer to archaea.

UR - http://www.scopus.com/inward/record.url?scp=70349667786&partnerID=8YFLogxK

U2 - 10.1128/EC.00161-09

DO - 10.1128/EC.00161-09

M3 - Article

VL - 8

SP - 1592

EP - 1603

JO - Eukaryotic Cell

JF - Eukaryotic Cell

SN - 1535-9778

IS - 10

ER -