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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.

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The expression of L. vannamei AMPs after dsLvLRRFIP2 was knocked down.The relative expression of the L. vannamei AMP genes ((A) LvPEN2, (B) LvPEN4, (C) LvCrustin, (D) LvALF1, (E) LvLyz1, (F) LvLyz2) were compared against the PBS and dsEGFP injection group at the corresponding times. Relative expression values were normalized to LvEF1α. The results are based on three independent experiments and expressed as mean values±SD. Statistical significance was calculated by the Student’s t-test (*indicates p<0.05 and **indicates p<0.01 compared with EGFP dsRNA injection group.
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pone-0057456-g008: The expression of L. vannamei AMPs after dsLvLRRFIP2 was knocked down.The relative expression of the L. vannamei AMP genes ((A) LvPEN2, (B) LvPEN4, (C) LvCrustin, (D) LvALF1, (E) LvLyz1, (F) LvLyz2) were compared against the PBS and dsEGFP injection group at the corresponding times. Relative expression values were normalized to LvEF1α. The results are based on three independent experiments and expressed as mean values±SD. Statistical significance was calculated by the Student’s t-test (*indicates p<0.05 and **indicates p<0.01 compared with EGFP dsRNA injection group.

Mentions: Considering that the knockdown of LvLRRFIP2 led to significantly increased mortality after V. parahaemolyticus infection (Fig. 7B), the expressions of six AMP genes were observed in LvLRRFIP2 knockdown L. vannamei. Fig. 8 shows that LvPEN4 underwent a brief period of downregulation at 0.5 d, 1 d, and 2 d after dsLvLRRFIP2 injection. However, the expression level of LvPEN4 was not significantly different in the dsLvLRRFIP2 and dsEGFP injection groups (Fig. 8B). Compared with the dsEGFP injection group, the expression level of LvPEN2, LvCrustin, LvALF1, LvLyz1, and LvLyz2 decreased at all detected times in the dsLvLRRFIP2 injection group (Figs. 8A, 8C, 8D, 8E, and 8F, respectively). All the results corresponded to the increase of the cumulative mortality of L. vannamei in the LvLRRFIP2 dsRNA group challenged with V. parahaemolyticus.


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)

The expression of L. vannamei AMPs after dsLvLRRFIP2 was knocked down.The relative expression of the L. vannamei AMP genes ((A) LvPEN2, (B) LvPEN4, (C) LvCrustin, (D) LvALF1, (E) LvLyz1, (F) LvLyz2) were compared against the PBS and dsEGFP injection group at the corresponding times. Relative expression values were normalized to LvEF1α. The results are based on three independent experiments and expressed as mean values±SD. Statistical significance was calculated by the Student’s t-test (*indicates p<0.05 and **indicates p<0.01 compared with EGFP dsRNA injection group.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0057456-g008: The expression of L. vannamei AMPs after dsLvLRRFIP2 was knocked down.The relative expression of the L. vannamei AMP genes ((A) LvPEN2, (B) LvPEN4, (C) LvCrustin, (D) LvALF1, (E) LvLyz1, (F) LvLyz2) were compared against the PBS and dsEGFP injection group at the corresponding times. Relative expression values were normalized to LvEF1α. The results are based on three independent experiments and expressed as mean values±SD. Statistical significance was calculated by the Student’s t-test (*indicates p<0.05 and **indicates p<0.01 compared with EGFP dsRNA injection group.
Mentions: Considering that the knockdown of LvLRRFIP2 led to significantly increased mortality after V. parahaemolyticus infection (Fig. 7B), the expressions of six AMP genes were observed in LvLRRFIP2 knockdown L. vannamei. Fig. 8 shows that LvPEN4 underwent a brief period of downregulation at 0.5 d, 1 d, and 2 d after dsLvLRRFIP2 injection. However, the expression level of LvPEN4 was not significantly different in the dsLvLRRFIP2 and dsEGFP injection groups (Fig. 8B). Compared with the dsEGFP injection group, the expression level of LvPEN2, LvCrustin, LvALF1, LvLyz1, and LvLyz2 decreased at all detected times in the dsLvLRRFIP2 injection group (Figs. 8A, 8C, 8D, 8E, and 8F, respectively). All the results corresponded to the increase of the cumulative mortality of L. vannamei in the LvLRRFIP2 dsRNA group challenged with V. parahaemolyticus.

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