Complete sequencing and expression of three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

Three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway have been fully sequenced and their expression investigated in rainbow trout (Oncorhynchus mykiss). Trout C1r cDNA encodes a 707-amino-acid (aa) protein with a theoretical M(r) of 77,200. The trout translation shows highest homology with carp C1r/s, and lower, equal homologies to mammalian C1r and C1s, and MASPs from other vertebrate species. However, phylogenetic analysis and structural features suggest that the trout sequence, together with the two carp sequences, are the orthologues of mammalian C1r. The trout C4 cDNA encodes a 1,724-aa protein with a theoretical M(r) of 192,600. The trout translation shows higher homologies to the carp C4B and medaka C4, but lower homologies to C4 from other species and the carp C4A. It has a predicted signal peptide of 22 aa, a alpha-chain of 773 aa, a beta-chain of 635 aa and a lambda-chain of 288 aa. Trout C1 inhibitor cDNA encodes a 611-aa protein with a theoretical M(r) of 68,700. The trout translation has a C-terminal serpin domain with high homologies with mammalian counterparts (~37% identities), and a longer N-terminus, with no significant homology to other serpins, which contains two Ig-like domains. A molecule containing two Ig-like domains followed by a serpin domain, has also been found in an EST clone from another bony fish, the Japanese flounder. This suggests a unique structural feature of C1 inhibitor in fish. The functional significance of the Ig domains is discussed. The liver is the major site of expression of the three trout complement components, C1r, C4 and C1 inhibitor, although their expression is also detectable in other tissues. The extra-hepatic expression of complement genes may be important for local protection and inflammatory responses. Low-level constitutive expression of the three components was also detectable in a trout monocyte/macrophage cell line RTS-11, but only the expression of C4 could be upregulated by LPS.
Original languageEnglish
Pages (from-to)615-628
Number of pages14
JournalImmunogenetics
Volume55
Issue number9
Early online date20 Nov 2003
DOIs
Publication statusPublished - Dec 2003

Keywords

  • Amino Acid Sequence
  • Animals
  • Cells, Cultured
  • Cloning, Molecular
  • Complement C1 Inactivator Proteins
  • Complement C1r
  • Complement C4
  • Complement Pathway, Classical
  • DNA, Complementary
  • Lipopolysaccharides
  • Molecular Sequence Data
  • Oncorhynchus mykiss
  • Phylogeny
  • Polymerase Chain Reaction
  • RNA
  • Sequence Homology, Amino Acid
  • Tissue Distribution
  • Rainbow Trout
  • Complement
  • C1r
  • C1 inhibitor
  • C4
  • Carp Cyprinus-Carpio
  • Molecular-Cloning
  • Serine-Protease
  • Reactive-Site
  • CDNA Cloning
  • 4th Component
  • Aeromonas-Salmonicida
  • Linkage Analysis
  • Multiple Forms
  • Serpin Domain

Cite this

@article{9b543a97ab1d4eab87b71be6e24a275d,
title = "Complete sequencing and expression of three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss",
abstract = "Three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway have been fully sequenced and their expression investigated in rainbow trout (Oncorhynchus mykiss). Trout C1r cDNA encodes a 707-amino-acid (aa) protein with a theoretical M(r) of 77,200. The trout translation shows highest homology with carp C1r/s, and lower, equal homologies to mammalian C1r and C1s, and MASPs from other vertebrate species. However, phylogenetic analysis and structural features suggest that the trout sequence, together with the two carp sequences, are the orthologues of mammalian C1r. The trout C4 cDNA encodes a 1,724-aa protein with a theoretical M(r) of 192,600. The trout translation shows higher homologies to the carp C4B and medaka C4, but lower homologies to C4 from other species and the carp C4A. It has a predicted signal peptide of 22 aa, a alpha-chain of 773 aa, a beta-chain of 635 aa and a lambda-chain of 288 aa. Trout C1 inhibitor cDNA encodes a 611-aa protein with a theoretical M(r) of 68,700. The trout translation has a C-terminal serpin domain with high homologies with mammalian counterparts (~37{\%} identities), and a longer N-terminus, with no significant homology to other serpins, which contains two Ig-like domains. A molecule containing two Ig-like domains followed by a serpin domain, has also been found in an EST clone from another bony fish, the Japanese flounder. This suggests a unique structural feature of C1 inhibitor in fish. The functional significance of the Ig domains is discussed. The liver is the major site of expression of the three trout complement components, C1r, C4 and C1 inhibitor, although their expression is also detectable in other tissues. The extra-hepatic expression of complement genes may be important for local protection and inflammatory responses. Low-level constitutive expression of the three components was also detectable in a trout monocyte/macrophage cell line RTS-11, but only the expression of C4 could be upregulated by LPS.",
keywords = "Amino Acid Sequence, Animals, Cells, Cultured, Cloning, Molecular, Complement C1 Inactivator Proteins, Complement C1r, Complement C4, Complement Pathway, Classical, DNA, Complementary, Lipopolysaccharides, Molecular Sequence Data, Oncorhynchus mykiss, Phylogeny, Polymerase Chain Reaction, RNA, Sequence Homology, Amino Acid, Tissue Distribution, Rainbow Trout, Complement , C1r, C1 inhibitor, C4, Carp Cyprinus-Carpio, Molecular-Cloning, Serine-Protease, Reactive-Site, CDNA Cloning, 4th Component , Aeromonas-Salmonicida , Linkage Analysis, Multiple Forms, Serpin Domain",
author = "Tiehui Wang and Secombes, {Christopher J}",
year = "2003",
month = "12",
doi = "10.1007/s00251-003-0622-5",
language = "English",
volume = "55",
pages = "615--628",
journal = "Immunogenetics",
issn = "0093-7711",
publisher = "Springer Verlag",
number = "9",

}

TY - JOUR

T1 - Complete sequencing and expression of three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss

AU - Wang, Tiehui

AU - Secombes, Christopher J

PY - 2003/12

Y1 - 2003/12

N2 - Three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway have been fully sequenced and their expression investigated in rainbow trout (Oncorhynchus mykiss). Trout C1r cDNA encodes a 707-amino-acid (aa) protein with a theoretical M(r) of 77,200. The trout translation shows highest homology with carp C1r/s, and lower, equal homologies to mammalian C1r and C1s, and MASPs from other vertebrate species. However, phylogenetic analysis and structural features suggest that the trout sequence, together with the two carp sequences, are the orthologues of mammalian C1r. The trout C4 cDNA encodes a 1,724-aa protein with a theoretical M(r) of 192,600. The trout translation shows higher homologies to the carp C4B and medaka C4, but lower homologies to C4 from other species and the carp C4A. It has a predicted signal peptide of 22 aa, a alpha-chain of 773 aa, a beta-chain of 635 aa and a lambda-chain of 288 aa. Trout C1 inhibitor cDNA encodes a 611-aa protein with a theoretical M(r) of 68,700. The trout translation has a C-terminal serpin domain with high homologies with mammalian counterparts (~37% identities), and a longer N-terminus, with no significant homology to other serpins, which contains two Ig-like domains. A molecule containing two Ig-like domains followed by a serpin domain, has also been found in an EST clone from another bony fish, the Japanese flounder. This suggests a unique structural feature of C1 inhibitor in fish. The functional significance of the Ig domains is discussed. The liver is the major site of expression of the three trout complement components, C1r, C4 and C1 inhibitor, although their expression is also detectable in other tissues. The extra-hepatic expression of complement genes may be important for local protection and inflammatory responses. Low-level constitutive expression of the three components was also detectable in a trout monocyte/macrophage cell line RTS-11, but only the expression of C4 could be upregulated by LPS.

AB - Three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway have been fully sequenced and their expression investigated in rainbow trout (Oncorhynchus mykiss). Trout C1r cDNA encodes a 707-amino-acid (aa) protein with a theoretical M(r) of 77,200. The trout translation shows highest homology with carp C1r/s, and lower, equal homologies to mammalian C1r and C1s, and MASPs from other vertebrate species. However, phylogenetic analysis and structural features suggest that the trout sequence, together with the two carp sequences, are the orthologues of mammalian C1r. The trout C4 cDNA encodes a 1,724-aa protein with a theoretical M(r) of 192,600. The trout translation shows higher homologies to the carp C4B and medaka C4, but lower homologies to C4 from other species and the carp C4A. It has a predicted signal peptide of 22 aa, a alpha-chain of 773 aa, a beta-chain of 635 aa and a lambda-chain of 288 aa. Trout C1 inhibitor cDNA encodes a 611-aa protein with a theoretical M(r) of 68,700. The trout translation has a C-terminal serpin domain with high homologies with mammalian counterparts (~37% identities), and a longer N-terminus, with no significant homology to other serpins, which contains two Ig-like domains. A molecule containing two Ig-like domains followed by a serpin domain, has also been found in an EST clone from another bony fish, the Japanese flounder. This suggests a unique structural feature of C1 inhibitor in fish. The functional significance of the Ig domains is discussed. The liver is the major site of expression of the three trout complement components, C1r, C4 and C1 inhibitor, although their expression is also detectable in other tissues. The extra-hepatic expression of complement genes may be important for local protection and inflammatory responses. Low-level constitutive expression of the three components was also detectable in a trout monocyte/macrophage cell line RTS-11, but only the expression of C4 could be upregulated by LPS.

KW - Amino Acid Sequence

KW - Animals

KW - Cells, Cultured

KW - Cloning, Molecular

KW - Complement C1 Inactivator Proteins

KW - Complement C1r

KW - Complement C4

KW - Complement Pathway, Classical

KW - DNA, Complementary

KW - Lipopolysaccharides

KW - Molecular Sequence Data

KW - Oncorhynchus mykiss

KW - Phylogeny

KW - Polymerase Chain Reaction

KW - RNA

KW - Sequence Homology, Amino Acid

KW - Tissue Distribution

KW - Rainbow Trout

KW - Complement

KW - C1r

KW - C1 inhibitor

KW - C4

KW - Carp Cyprinus-Carpio

KW - Molecular-Cloning

KW - Serine-Protease

KW - Reactive-Site

KW - CDNA Cloning

KW - 4th Component

KW - Aeromonas-Salmonicida

KW - Linkage Analysis

KW - Multiple Forms

KW - Serpin Domain

U2 - 10.1007/s00251-003-0622-5

DO - 10.1007/s00251-003-0622-5

M3 - Article

VL - 55

SP - 615

EP - 628

JO - Immunogenetics

JF - Immunogenetics

SN - 0093-7711

IS - 9

ER -