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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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Gene silencing of L. vannamei LvLRRFIP2 increased its mortality after V. parahaemolyticus injection.L. vannamei were injected intramuscularly with PBS or dsRNAs corresponding to LvLRRFIP2 or EGFP. At 2 d after the initial injection, the animals were infected with PBS (negative control) (A), V. parahaemolyticus (B), S. aureus (C), or WSSV (D). Mortality was measured in each treatment group (n = 50) and was recorded every 8 h post-challenge. Differences in cumulative mortality levels between the LvLRRFIP2 dsRNA group and the EGFP dsRNA group were analyzed by Kaplan-Meier log-rank χ2 tests. Significant differences in L. vannamei mortality are marked with asterisks, and were found only in L. vannamei challenged with V. parahaemolyticus from 88 hpi to the end of the experiment (p<0.05).
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pone-0057456-g007: Gene silencing of L. vannamei LvLRRFIP2 increased its mortality after V. parahaemolyticus injection.L. vannamei were injected intramuscularly with PBS or dsRNAs corresponding to LvLRRFIP2 or EGFP. At 2 d after the initial injection, the animals were infected with PBS (negative control) (A), V. parahaemolyticus (B), S. aureus (C), or WSSV (D). Mortality was measured in each treatment group (n = 50) and was recorded every 8 h post-challenge. Differences in cumulative mortality levels between the LvLRRFIP2 dsRNA group and the EGFP dsRNA group were analyzed by Kaplan-Meier log-rank χ2 tests. Significant differences in L. vannamei mortality are marked with asterisks, and were found only in L. vannamei challenged with V. parahaemolyticus from 88 hpi to the end of the experiment (p<0.05).

Mentions: L. vannamei was challenged with V. parahaemolyticus, S. aureus, WSSV, and a PBS control to explore the possible involvement of LvLRRFIP2 in a protective response against invaders at 2 d post-dsRNA injection. The baseline cumulative mortality of L. vannamei injected with PBS at 2 d after LvLRRFIP2 dsRNA injection is shown in Fig. 7A. The final mortality rates at 136 hpi were low for all groups (11.6%, 9.3%, and 6.9% for the LvLRRFIP2 dsRNA, EGFP dsRNA, and PBS groups, respectively), and no significant difference in mortality was observed among the three groups (p>0.05). In the V. parahaemolyticus challenge test (Fig. 7B), the cumulative mortality of the LvLRRFIP2 dsRNA group began to increase at 8 h post-V. parahaemolyticus challenge. The cumulative mortality in the LvLRRFIP2 dsRNA group was significantly higher than in the EGFP dsRNA, starting at 88 hpi (Kaplan-Meier log-rank χ2: 7.402, p<0.05). The final mortality rates at 136 hpi were 63.6%, 30.0%, and 27.9% for the LvLRRFIP2 dsRNA, EGFP dsRNA, and PBS groups, respectively. In the challenge tests with S. aureus and WSSV, no evidence showed that knockdown of LvLRRFIP2 expression by dsRNA had any statistically significance on cumulative mortality (Figs. 7C and 7D).


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)

Gene silencing of L. vannamei LvLRRFIP2 increased its mortality after V. parahaemolyticus injection.L. vannamei were injected intramuscularly with PBS or dsRNAs corresponding to LvLRRFIP2 or EGFP. At 2 d after the initial injection, the animals were infected with PBS (negative control) (A), V. parahaemolyticus (B), S. aureus (C), or WSSV (D). Mortality was measured in each treatment group (n = 50) and was recorded every 8 h post-challenge. Differences in cumulative mortality levels between the LvLRRFIP2 dsRNA group and the EGFP dsRNA group were analyzed by Kaplan-Meier log-rank χ2 tests. Significant differences in L. vannamei mortality are marked with asterisks, and were found only in L. vannamei challenged with V. parahaemolyticus from 88 hpi to the end of the experiment (p<0.05).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585381&req=5

pone-0057456-g007: Gene silencing of L. vannamei LvLRRFIP2 increased its mortality after V. parahaemolyticus injection.L. vannamei were injected intramuscularly with PBS or dsRNAs corresponding to LvLRRFIP2 or EGFP. At 2 d after the initial injection, the animals were infected with PBS (negative control) (A), V. parahaemolyticus (B), S. aureus (C), or WSSV (D). Mortality was measured in each treatment group (n = 50) and was recorded every 8 h post-challenge. Differences in cumulative mortality levels between the LvLRRFIP2 dsRNA group and the EGFP dsRNA group were analyzed by Kaplan-Meier log-rank χ2 tests. Significant differences in L. vannamei mortality are marked with asterisks, and were found only in L. vannamei challenged with V. parahaemolyticus from 88 hpi to the end of the experiment (p<0.05).
Mentions: L. vannamei was challenged with V. parahaemolyticus, S. aureus, WSSV, and a PBS control to explore the possible involvement of LvLRRFIP2 in a protective response against invaders at 2 d post-dsRNA injection. The baseline cumulative mortality of L. vannamei injected with PBS at 2 d after LvLRRFIP2 dsRNA injection is shown in Fig. 7A. The final mortality rates at 136 hpi were low for all groups (11.6%, 9.3%, and 6.9% for the LvLRRFIP2 dsRNA, EGFP dsRNA, and PBS groups, respectively), and no significant difference in mortality was observed among the three groups (p>0.05). In the V. parahaemolyticus challenge test (Fig. 7B), the cumulative mortality of the LvLRRFIP2 dsRNA group began to increase at 8 h post-V. parahaemolyticus challenge. The cumulative mortality in the LvLRRFIP2 dsRNA group was significantly higher than in the EGFP dsRNA, starting at 88 hpi (Kaplan-Meier log-rank χ2: 7.402, p<0.05). The final mortality rates at 136 hpi were 63.6%, 30.0%, and 27.9% for the LvLRRFIP2 dsRNA, EGFP dsRNA, and PBS groups, respectively. In the challenge tests with S. aureus and WSSV, no evidence showed that knockdown of LvLRRFIP2 expression by dsRNA had any statistically significance on cumulative mortality (Figs. 7C and 7D).

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