Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA

Tafalla, Carolina; Aranguren, Raquel; Secombes, Christopher J.; Figueras, António; Novoa, Beatriz

doi:10.1016/s1050-4648(03)00010-x

Cited by 49 publications

(44 citation statements)

References 23 publications

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“…Since then, Mx genes have been identified in a variety of mammals (Horisberger and Gunst 1991), birds (Bazzigher et al 1993;Ko et al 2002), and teleost fish (Trobridge and Leong 1995;Robertsen et al 1997;Jensen and Robertsen 2000;Lee et al 2000;Yap et al 2003;Tafalla et al 2004;Altmann et al 2004). They are large GTPases that belong to the dynamin superfamily (Leong et al 1998).…”

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confidence: 99%

“…The amino-acid residues identical to those of CaMx1 identified by the CLUSTAL W program (Thompson et al 1994) are indicated by asterisks; dashes indicate gaps introduced for optimal alignment. Ca Crucain carp, Dr zebrafish, Om rainbow trout, Ss Atlantic salmon, Hs human, Gg chicken system gene, Mx has been characterized from many fish species: seven copies for zebrafish (Lutfalla et al 2003), three copies for rainbow trout (Trobridge and Leong 1995;Trobridge et al 1997) and Atlantic salmon (Robertsen et al 1997); a single copy for perch (Staeheli et al 1989), Japanese flounder (Lee et al 2000), Atlantic halibut (Jensen and Robertsen 2000), gilthead sea bream (Tafalla et al 2004) and fugu (Yap et al 2003). The fugu Mx gene organization and regulatory region have also been characterized (Yap et al 2003).…”

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confidence: 99%

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Differential expression of two Carassius auratus Mx genes in cultured CAB cells induced by grass carp hemorrhage virus and interferon

Zhang

Gui

2004

Immunogenetics

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UV-inactivated GCHV (grass carp hemorrhage virus) is able to induce an antiviral state in cultured CAB cells (crucian carp Carassius auratus blastulae embryonic cells) via the production of interferon (IFN). In the current work, the full-length cDNAs of two Mx genes, termed CaMx1 and CaMx2, have been cloned and sequenced from UV-inactivated GCHV-infected and still IFN-producing CAB cells by suppression subtractive hybridization. Their putative proteins show the characteristically structural features of mammalian IFN-induced Mx proteins, including GTP-binding motif, dynamin family signature and leucine zipper motif. CaMx1 exhibits 85% sequence identity to zebrafish MxA and 72-74% to three Atlantic salmon Mx proteins. CaMx2 is most similar to zebrafish MxE, with 80% identity, and then rainbow trout Mx3, with 52%. Constitutive expression was detected by RT-PCR for CaMx1, but not for CaMx2, in normal CAB cells, but their up-regulations could be induced after treatment with active GCHV, UV-inactivated GCHV and CAB IFN. Distinct kinetics of expression was observed for either CaMx1 or CaMx2 corresponding to the three stimuli, and even between CaMx1 and CaMx2, corresponding to the same stimulus. Upon virus infection, the transcriptional induction was strongly blocked for CaMx2 by cycloheximide (CHX), whereas almost nothing was observed for CaMx1. By contrast, following treatment with CAB IFN, CHX did not inhibit either gene transcription. Collectively, these results suggest that there are very distinct mechanisms for modulating the expression of both CaMx1 and CaMx2 in normal and GCHV-infected CAB cells.

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confidence: 99%

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confidence: 99%

Differential expression of two Carassius auratus Mx genes in cultured CAB cells induced by grass carp hemorrhage virus and interferon

Zhang

Gui

2004

Immunogenetics

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“…Many kinds of fish species had been injected with poly I:C for induction of Mx gene expression [16e18,21e23,38]. In vitro, Mx gene expression could also be induced by poly I:C in some fish cell lines, such as RTG-2 cells [16], CHSE-214 [39], SAF-1 [21], and CCO cells [22]. In this study, cBB cells expressed Mx gene by poly I:C transfection either at 28 C or 20 C, but the expression level of Mx at 20 C was lower and delayed compared to that at 28 C, which was an optimal culture temperature for barramundi.…”

Section: Discussionmentioning

confidence: 99%

“…Mx induction by fish viruses has been reported in many cases, including channel catfish virus (CCV) in catfish [22], nodavirus and iridovirus in grouper [23], infectious hematopoietic necrosis virus (IHNV) in rainbow trout [16], IPNV in Atlantic halibut [18], hirame rhabdovirus (HIRRV) or aquabirnavirus in Japanese flounder [19,40] in vivo, and nodavirus in SAF-1 cells [21] in vitro. In the present study, we found that cBB cells were sensitive to nodavirus, birnavirus and iridovirus infection.…”

Section: Discussionmentioning

confidence: 99%

“…In fish, the first Mx gene was identified from perch (Perca fluviatilis L.) [15]. Following, piscine Mx genes have been cloned and characterized in rainbow trout (Oncorhynchus mykiss Walbaum) [16], Atlantic salmon (Salmo salar L.) [17], Atlantic halibut (Hippoglossus hippoglossus L.) [18], Japanese flounder (Paralichthys olivaceus) [19], fugu (Takifugu rubripes) [20], gilthead sea bream (Sparus aurata) [21], channel catfish (Ictalurus punctatus) [22], and orange-spotted grouper (Epinephelus coioides) [23]. However, only few papers have demonstrated the role of fish Mx proteins in IFN response.…”

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confidence: 99%

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Cloning and analysis of antiviral activity of a barramundi (Lates calcarifer) Mx gene☆

Chi

2007

Fish & Shellfish Immunology

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We obtained a full-length cDNA clone for the Mx gene of barramundi (Lates calcarifer), using RACE (rapid amplification of cDNA ends) polymerase chain reaction ( C was higher than that at 20 C. Moreover, Mx gene expression was also induced by viral infection, including fish nodavirus, birnavirus, and iridovirus. Among these, nodavirus was a stronger inducer than the other two viruses. Using an antiviral activity assay, we revealed that poly I:C-transfected cBB cells had antiviral activity against fish nodavirus and birnavirus, but not iridovirus. Furthermore, the replication of nodavirus and birnavirus could be restored after the expression of Mx gene was down-regulated by siRNA. Therefore, these results indicated that the expression of barramundi Mx gene was able to inhibit the proliferation of fish nodavirus and birnavirus.

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The piscine SAF-1 cell line: genetic stability and labeling

et al. 2005

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Fish cell lines are increasingly important research tools. The SAF-1 cell line, fibroblast-like culture derived from the marine fish gilthead seabream (Sparus aurata), has proved useful in many applications, especially in viral research. For cell lines intended as in vitro models, characterization of their properties and authentication are essential for deeper understanding of their performance and thus more precise experimental design and applicability. In this study we characterized the SAF-1 cell line in terms of genetic stability through time and genetic labeling. Methods for determining stability include telomerase activity, karyotyping, mapping of ribosomal RNA regions, and DNA content. For genetic labeling 12 microsatellite loci were used. The results indicate that telomerase has been activated in the course of SAF-1 development, and the highest levels of telomerase activity correlate with an increase in cell proliferation, thus supporting a permanent cell line. This stability is in agreement with the normal situation presented by the cytogenetic traits and DNA content values, and the genotypic profile allows SAF-1 authentication at the single individual level. This study increases the value of SAF-1 as an in vitro system, which is now one of the few well-characterized cell lines from a marine fish.

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Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA

Cited by 49 publications

References 23 publications

Differential expression of two Carassius auratus Mx genes in cultured CAB cells induced by grass carp hemorrhage virus and interferon

Differential expression of two Carassius auratus Mx genes in cultured CAB cells induced by grass carp hemorrhage virus and interferon

Cloning and analysis of antiviral activity of a barramundi (Lates calcarifer) Mx gene☆

The piscine SAF-1 cell line: genetic stability and labeling

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