A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva

As a first step in developing compartment-specific markers for protein trafficking within Theileria parva, we have isolated cDNAs encoding homologues of the small GTP binding proteins Rab1 and Rab4. The T. parva homologue of Rab1 (TpRab1), a protein which regulates vesicular transport between the en...

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Autores principales: Janoo, R.T.K., Musoke, A.J., Wells, C., Bishop, Richard P.
Formato: Journal Article
Lenguaje:Inglés
Publicado: Elsevier 1999
Materias:
Acceso en línea:https://hdl.handle.net/10568/33063
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author Janoo, R.T.K.
Musoke, A.J.
Wells, C.
Bishop, Richard P.
author_browse Bishop, Richard P.
Janoo, R.T.K.
Musoke, A.J.
Wells, C.
author_facet Janoo, R.T.K.
Musoke, A.J.
Wells, C.
Bishop, Richard P.
author_sort Janoo, R.T.K.
collection Repository of Agricultural Research Outputs (CGSpace)
description As a first step in developing compartment-specific markers for protein trafficking within Theileria parva, we have isolated cDNAs encoding homologues of the small GTP binding proteins Rab1 and Rab4. The T. parva homologue of Rab1 (TpRab1), a protein which regulates vesicular transport between the endoplasmic reticulum and cis golgi in other organisms, was unusual in that it contained a unique 17 amino acid C-terminal extension. The C-terminal motifisequence KCT (XCX) contrasted with the CXC or XCC motifs which act as signals for isoprenylation by geranylgeranyl in most Rab proteins, including all known Rab1 homologues, in containing only a single cysteine, (C14) mevalonic acid lactone and (H3) geranylgeranyl pyrophosphate were specifically incorporated into recombinant TpRab1 in vitro, demonstrating that the novel motif was functional for isoprenylation. Recombinant TpRab1 bound radiolabeled GTP, and this binding was inhibited by excess unlabeled GTP and GDP and also partially by ATP. The TpRab1 gene contained four short (34-67 bp) introns with a distinct pattern of occurrence within the protein sequence as compared to the introns of other lower eukaryote Rab1 genes. Immunofluorescence microscopy using antiserum specific for the novel C-terminal peptide in combination with labelling of cells using the nucleic acid-staining dye DAPI, indicated that TpRab1 was located in the vicinity of the schizont nucleus within the infected lymphocyte.
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spelling CGSpace330632024-05-01T08:18:11Z A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva Janoo, R.T.K. Musoke, A.J. Wells, C. Bishop, Richard P. theileria parva dna genetic markers lymphocytes As a first step in developing compartment-specific markers for protein trafficking within Theileria parva, we have isolated cDNAs encoding homologues of the small GTP binding proteins Rab1 and Rab4. The T. parva homologue of Rab1 (TpRab1), a protein which regulates vesicular transport between the endoplasmic reticulum and cis golgi in other organisms, was unusual in that it contained a unique 17 amino acid C-terminal extension. The C-terminal motifisequence KCT (XCX) contrasted with the CXC or XCC motifs which act as signals for isoprenylation by geranylgeranyl in most Rab proteins, including all known Rab1 homologues, in containing only a single cysteine, (C14) mevalonic acid lactone and (H3) geranylgeranyl pyrophosphate were specifically incorporated into recombinant TpRab1 in vitro, demonstrating that the novel motif was functional for isoprenylation. Recombinant TpRab1 bound radiolabeled GTP, and this binding was inhibited by excess unlabeled GTP and GDP and also partially by ATP. The TpRab1 gene contained four short (34-67 bp) introns with a distinct pattern of occurrence within the protein sequence as compared to the introns of other lower eukaryote Rab1 genes. Immunofluorescence microscopy using antiserum specific for the novel C-terminal peptide in combination with labelling of cells using the nucleic acid-staining dye DAPI, indicated that TpRab1 was located in the vicinity of the schizont nucleus within the infected lymphocyte. 1999-07 2013-07-03T05:26:00Z 2013-07-03T05:26:00Z Journal Article https://hdl.handle.net/10568/33063 en Limited Access Elsevier Molecular and Biochemical Parasitology;102: 131-143
spellingShingle theileria parva
dna
genetic markers
lymphocytes
Janoo, R.T.K.
Musoke, A.J.
Wells, C.
Bishop, Richard P.
A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title_full A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title_fullStr A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title_full_unstemmed A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title_short A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva
title_sort rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of theileria parva
topic theileria parva
dna
genetic markers
lymphocytes
url https://hdl.handle.net/10568/33063
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