Limits...
Identification and function of leucine-rich repeat flightless-I-interacting protein 2 (LRRFIP2) in Litopenaeus vannamei.

Zhang S, Yan H, Li CZ, Chen YH, Yuan FH, Chen YG, Weng SP, He JG - PLoS ONE (2013)

Bottom Line: The knockdown of LvLRRFIP2 by RNA interference resulted in higher cumulative mortality of L. vannamei upon V. parahaemolyticus but not S. aureus and WSSV infections.The expression of L. vannamei AMP genes were reduced by dsLvLRRFIP2 interference.These results indicate that LvLRRFIP2 has an important function in antibacterials via the regulation of AMP gene expression.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory of Aquatic Product Safety/State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, People's Republic of China.

ABSTRACT
Leucine-rich repeat flightless-I-interacting protein 2 (LRRFIP2) is a myeloid differentiation factor 88-interacting protein with a positive regulatory function in toll-like receptor signaling. In this study, seven LRRFIP2 protein variants (LvLRRFIP2A-G) were identified in Litopenaeus vannamei. All the seven LvLRRFIP2 protein variants encode proteins with a DUF2051 domain. LvLRRFIP2s were upregulated in hemocytes after challenged with lipopolysaccharide, poly I:C, CpG-ODN2006, Vibrio parahaemolyticus, Staphylococcus aureus, and white spot syndrome virus (WSSV). Dual-luciferase reporter assays in Drosophila Schneider 2 cells revealed that LvLRRFIP2 activates the promoters of Drosophila and shrimp AMP genes. The knockdown of LvLRRFIP2 by RNA interference resulted in higher cumulative mortality of L. vannamei upon V. parahaemolyticus but not S. aureus and WSSV infections. The expression of L. vannamei AMP genes were reduced by dsLvLRRFIP2 interference. These results indicate that LvLRRFIP2 has an important function in antibacterials via the regulation of AMP gene expression.

Show MeSH

Related in: MedlinePlus

Effects of LvLRRFIP2s on the promoter activities of Drosophila and shrimp AMPs in Drosophila S2 cells.Drosophila S2 cells were transfected with the protein expression vector (pAC5.1 empty vector, one of the LvLRRFIP2s), the reporter gene plasmid (pGL3-Basic, pGL3-AttA, pGL3-Drs, pGL3-PEN453, or pGL3-PEN536), and the pRL-TK Renilla luciferase plasmid (as an internal control: Promega, USA). After 48 h, the cells were harvested for luciferase activity determination using the dual-luciferase reporter assay system (Promega, USA). All data are representative of three independent experiments. The bars indicate the mean ± SD of luciferase activity (n = 3). The statistical significance was calculated using Student’s t-test (*indicates p<0.05 and **indicates p<0.01compared with control).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3585381&req=5

pone-0057456-g005: Effects of LvLRRFIP2s on the promoter activities of Drosophila and shrimp AMPs in Drosophila S2 cells.Drosophila S2 cells were transfected with the protein expression vector (pAC5.1 empty vector, one of the LvLRRFIP2s), the reporter gene plasmid (pGL3-Basic, pGL3-AttA, pGL3-Drs, pGL3-PEN453, or pGL3-PEN536), and the pRL-TK Renilla luciferase plasmid (as an internal control: Promega, USA). After 48 h, the cells were harvested for luciferase activity determination using the dual-luciferase reporter assay system (Promega, USA). All data are representative of three independent experiments. The bars indicate the mean ± SD of luciferase activity (n = 3). The statistical significance was calculated using Student’s t-test (*indicates p<0.05 and **indicates p<0.01compared with control).

Mentions: LRRFIP2 is a positive regulator of NF-κB activity in murine macrophage cells [8]. AMPs are important immune factors in Drosophila and shrimp, and their expression is believed to be controlled mainly by the NF-κB signal pathway [20], [21], [32]. The NF-κB signal pathway can be activated by Toll, Pelle, TRAF6, Dorsal, and Relish in shrimp [19]–[24]. The present study demonstrated that LvLRRFIP2s activated the promoters of Drosophila and shrimp AMP genes. Compared with six other variants of LvLRRFIP2, LvLRRFIP2F induced higher activities of AMP promoters, including the Drosophila AMPs AttA (3.78-fold), Drs (2.11-fold), L. vannamei AMP PEN4 (3.32-fold), and P. monodon AMP PEN536 (5.14-fold) (Fig. 5).


Identification and function of leucine-rich repeat flightless-I-interacting protein 2 (LRRFIP2) in Litopenaeus vannamei.

Zhang S, Yan H, Li CZ, Chen YH, Yuan FH, Chen YG, Weng SP, He JG - PLoS ONE (2013)

Effects of LvLRRFIP2s on the promoter activities of Drosophila and shrimp AMPs in Drosophila S2 cells.Drosophila S2 cells were transfected with the protein expression vector (pAC5.1 empty vector, one of the LvLRRFIP2s), the reporter gene plasmid (pGL3-Basic, pGL3-AttA, pGL3-Drs, pGL3-PEN453, or pGL3-PEN536), and the pRL-TK Renilla luciferase plasmid (as an internal control: Promega, USA). After 48 h, the cells were harvested for luciferase activity determination using the dual-luciferase reporter assay system (Promega, USA). All data are representative of three independent experiments. The bars indicate the mean ± SD of luciferase activity (n = 3). The statistical significance was calculated using Student’s t-test (*indicates p<0.05 and **indicates p<0.01compared with control).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585381&req=5

pone-0057456-g005: Effects of LvLRRFIP2s on the promoter activities of Drosophila and shrimp AMPs in Drosophila S2 cells.Drosophila S2 cells were transfected with the protein expression vector (pAC5.1 empty vector, one of the LvLRRFIP2s), the reporter gene plasmid (pGL3-Basic, pGL3-AttA, pGL3-Drs, pGL3-PEN453, or pGL3-PEN536), and the pRL-TK Renilla luciferase plasmid (as an internal control: Promega, USA). After 48 h, the cells were harvested for luciferase activity determination using the dual-luciferase reporter assay system (Promega, USA). All data are representative of three independent experiments. The bars indicate the mean ± SD of luciferase activity (n = 3). The statistical significance was calculated using Student’s t-test (*indicates p<0.05 and **indicates p<0.01compared with control).
Mentions: LRRFIP2 is a positive regulator of NF-κB activity in murine macrophage cells [8]. AMPs are important immune factors in Drosophila and shrimp, and their expression is believed to be controlled mainly by the NF-κB signal pathway [20], [21], [32]. The NF-κB signal pathway can be activated by Toll, Pelle, TRAF6, Dorsal, and Relish in shrimp [19]–[24]. The present study demonstrated that LvLRRFIP2s activated the promoters of Drosophila and shrimp AMP genes. Compared with six other variants of LvLRRFIP2, LvLRRFIP2F induced higher activities of AMP promoters, including the Drosophila AMPs AttA (3.78-fold), Drs (2.11-fold), L. vannamei AMP PEN4 (3.32-fold), and P. monodon AMP PEN536 (5.14-fold) (Fig. 5).

Bottom Line: The knockdown of LvLRRFIP2 by RNA interference resulted in higher cumulative mortality of L. vannamei upon V. parahaemolyticus but not S. aureus and WSSV infections.The expression of L. vannamei AMP genes were reduced by dsLvLRRFIP2 interference.These results indicate that LvLRRFIP2 has an important function in antibacterials via the regulation of AMP gene expression.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory of Aquatic Product Safety/State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, People's Republic of China.

ABSTRACT
Leucine-rich repeat flightless-I-interacting protein 2 (LRRFIP2) is a myeloid differentiation factor 88-interacting protein with a positive regulatory function in toll-like receptor signaling. In this study, seven LRRFIP2 protein variants (LvLRRFIP2A-G) were identified in Litopenaeus vannamei. All the seven LvLRRFIP2 protein variants encode proteins with a DUF2051 domain. LvLRRFIP2s were upregulated in hemocytes after challenged with lipopolysaccharide, poly I:C, CpG-ODN2006, Vibrio parahaemolyticus, Staphylococcus aureus, and white spot syndrome virus (WSSV). Dual-luciferase reporter assays in Drosophila Schneider 2 cells revealed that LvLRRFIP2 activates the promoters of Drosophila and shrimp AMP genes. The knockdown of LvLRRFIP2 by RNA interference resulted in higher cumulative mortality of L. vannamei upon V. parahaemolyticus but not S. aureus and WSSV infections. The expression of L. vannamei AMP genes were reduced by dsLvLRRFIP2 interference. These results indicate that LvLRRFIP2 has an important function in antibacterials via the regulation of AMP gene expression.

Show MeSH
Related in: MedlinePlus