The putative RxLR effector protein SpHtp1 from the fish pathogenic oomycete Saprolegnia parasitica is translocated into fish cells

Pieter van West, Irene de Bruijn, Kirsty L Minor, Andrew J Phillips, Emma J Robertson, Stephan Wawra, Judith Bain, Victoria L Anderson, Chris J Secombes

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The fish pathogenic oomycete Saprolegnia parasitica causes the disease Saprolegniosis in salmonids and other freshwater fish, resulting in considerable economic losses in aquaculture. Very little is known about the molecular and cellular mechanisms underlying the infection process of fish pathogenic oomycetes. In order to investigate the interaction in detail, an in vitro infection assay using an Oncorhynchus mykiss (rainbow trout) cell line (RTG-2) was developed. In a zoospore/cyst cDNA library, we identified the ORF SpHtp1, which encodes a secreted protein containing an RxLR motif. Detailed expression analysis indicated that SpHtp1 is highly expressed in zoospores/cysts from S. parasitica and in the very early stages of infection on RTG-2 cells, when compared with in vitro-grown mycelium. Moreover, the protein, SpHtp1, was found to translocate into the RTG-2 trout cells, during the interaction with S. parasitica, and also when the RTG-2 cells were treated with recombinant SpHtp1 fused to a C-terminal His-tag. These findings suggest that protein translocation could play an important role in Saprolegniosis.
Original languageEnglish
Pages (from-to)127-137
Number of pages11
JournalFEMS Microbiology Letters
Volume310
Issue number2
Early online date9 Jul 2010
DOIs
Publication statusPublished - Sep 2010

Fingerprint

Saprolegnia
Fish Proteins
Oomycetes
Fishes
Oncorhynchus mykiss
Cysts
Infection
Salmonidae
Aquaculture
Trout
Mycelium
Protein Transport
Fresh Water
Gene Library
Cell Communication
Open Reading Frames
Proteins
Economics
Cell Line
In Vitro Techniques

Keywords

  • amino acid sequence
  • animals
  • cell line
  • fish diseases
  • gene expression profiling
  • gene expression regulation
  • host-parasite interactions
  • infection
  • molecular sequence data
  • Oncorhynchus mykiss
  • protein transport
  • protozoan proteins
  • saprolegnia

Cite this

The putative RxLR effector protein SpHtp1 from the fish pathogenic oomycete Saprolegnia parasitica is translocated into fish cells. / van West, Pieter; de Bruijn, Irene; Minor, Kirsty L; Phillips, Andrew J; Robertson, Emma J; Wawra, Stephan; Bain, Judith; Anderson, Victoria L; Secombes, Chris J.

In: FEMS Microbiology Letters, Vol. 310, No. 2, 09.2010, p. 127-137.

Research output: Contribution to journalArticle

van West, Pieter ; de Bruijn, Irene ; Minor, Kirsty L ; Phillips, Andrew J ; Robertson, Emma J ; Wawra, Stephan ; Bain, Judith ; Anderson, Victoria L ; Secombes, Chris J. / The putative RxLR effector protein SpHtp1 from the fish pathogenic oomycete Saprolegnia parasitica is translocated into fish cells. In: FEMS Microbiology Letters. 2010 ; Vol. 310, No. 2. pp. 127-137.
@article{f7a69bee362d4cbc970451b0720aceed,
title = "The putative RxLR effector protein SpHtp1 from the fish pathogenic oomycete Saprolegnia parasitica is translocated into fish cells",
abstract = "The fish pathogenic oomycete Saprolegnia parasitica causes the disease Saprolegniosis in salmonids and other freshwater fish, resulting in considerable economic losses in aquaculture. Very little is known about the molecular and cellular mechanisms underlying the infection process of fish pathogenic oomycetes. In order to investigate the interaction in detail, an in vitro infection assay using an Oncorhynchus mykiss (rainbow trout) cell line (RTG-2) was developed. In a zoospore/cyst cDNA library, we identified the ORF SpHtp1, which encodes a secreted protein containing an RxLR motif. Detailed expression analysis indicated that SpHtp1 is highly expressed in zoospores/cysts from S. parasitica and in the very early stages of infection on RTG-2 cells, when compared with in vitro-grown mycelium. Moreover, the protein, SpHtp1, was found to translocate into the RTG-2 trout cells, during the interaction with S. parasitica, and also when the RTG-2 cells were treated with recombinant SpHtp1 fused to a C-terminal His-tag. These findings suggest that protein translocation could play an important role in Saprolegniosis.",
keywords = "amino acid sequence, animals, cell line, fish diseases, gene expression profiling, gene expression regulation, host-parasite interactions, infection, molecular sequence data , Oncorhynchus mykiss, protein transport, protozoan proteins, saprolegnia",
author = "{van West}, Pieter and {de Bruijn}, Irene and Minor, {Kirsty L} and Phillips, {Andrew J} and Robertson, {Emma J} and Stephan Wawra and Judith Bain and Anderson, {Victoria L} and Secombes, {Chris J}",
year = "2010",
month = "9",
doi = "10.1111/j.1574-6968.2010.02055.x",
language = "English",
volume = "310",
pages = "127--137",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - The putative RxLR effector protein SpHtp1 from the fish pathogenic oomycete Saprolegnia parasitica is translocated into fish cells

AU - van West, Pieter

AU - de Bruijn, Irene

AU - Minor, Kirsty L

AU - Phillips, Andrew J

AU - Robertson, Emma J

AU - Wawra, Stephan

AU - Bain, Judith

AU - Anderson, Victoria L

AU - Secombes, Chris J

PY - 2010/9

Y1 - 2010/9

N2 - The fish pathogenic oomycete Saprolegnia parasitica causes the disease Saprolegniosis in salmonids and other freshwater fish, resulting in considerable economic losses in aquaculture. Very little is known about the molecular and cellular mechanisms underlying the infection process of fish pathogenic oomycetes. In order to investigate the interaction in detail, an in vitro infection assay using an Oncorhynchus mykiss (rainbow trout) cell line (RTG-2) was developed. In a zoospore/cyst cDNA library, we identified the ORF SpHtp1, which encodes a secreted protein containing an RxLR motif. Detailed expression analysis indicated that SpHtp1 is highly expressed in zoospores/cysts from S. parasitica and in the very early stages of infection on RTG-2 cells, when compared with in vitro-grown mycelium. Moreover, the protein, SpHtp1, was found to translocate into the RTG-2 trout cells, during the interaction with S. parasitica, and also when the RTG-2 cells were treated with recombinant SpHtp1 fused to a C-terminal His-tag. These findings suggest that protein translocation could play an important role in Saprolegniosis.

AB - The fish pathogenic oomycete Saprolegnia parasitica causes the disease Saprolegniosis in salmonids and other freshwater fish, resulting in considerable economic losses in aquaculture. Very little is known about the molecular and cellular mechanisms underlying the infection process of fish pathogenic oomycetes. In order to investigate the interaction in detail, an in vitro infection assay using an Oncorhynchus mykiss (rainbow trout) cell line (RTG-2) was developed. In a zoospore/cyst cDNA library, we identified the ORF SpHtp1, which encodes a secreted protein containing an RxLR motif. Detailed expression analysis indicated that SpHtp1 is highly expressed in zoospores/cysts from S. parasitica and in the very early stages of infection on RTG-2 cells, when compared with in vitro-grown mycelium. Moreover, the protein, SpHtp1, was found to translocate into the RTG-2 trout cells, during the interaction with S. parasitica, and also when the RTG-2 cells were treated with recombinant SpHtp1 fused to a C-terminal His-tag. These findings suggest that protein translocation could play an important role in Saprolegniosis.

KW - amino acid sequence

KW - animals

KW - cell line

KW - fish diseases

KW - gene expression profiling

KW - gene expression regulation

KW - host-parasite interactions

KW - infection

KW - molecular sequence data

KW - Oncorhynchus mykiss

KW - protein transport

KW - protozoan proteins

KW - saprolegnia

U2 - 10.1111/j.1574-6968.2010.02055.x

DO - 10.1111/j.1574-6968.2010.02055.x

M3 - Article

VL - 310

SP - 127

EP - 137

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 2

ER -