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PLoS One ; 8(8): e72793, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24013515

RESUMO

Astakine is an important cytokine that is involved in crustacean hematopoiesis. Interestingly, the protein levels of astakine increased dramatically in plasma of LPS-injected shrimp while mRNA levels remained unchanged. Here, we investigated the involvement of astakine 3'-untranslated region (UTR) in its protein expression. The 3'-UTR of astakine down-regulated the expression of reporter protein but the mRNA stability of reporter gene was unaffected. We identified the functional regulatory elements of astakine 3'-UTR, where 3'-UTR242-483 acted as repressor. The electrophoresis mobility shift assay (EMSA), RNA pull-down assay and LC/MS/MS were performed to identify the protein association. We noted that crustin Pm4 and shrimp transglutaminase I (STG I) were associated to astakine 3'-UTR242-483, while two other proteins have yet to be revealed. Depletion of hemocytic crustin Pm4 and STG I significantly increased the protein level of astakine while astakine mRNA level remained unaffected. Lipopolysaccharide (LPS) stimulated the secretion of crustin Pm4 and STG I from hemocytes to plasma and increased the astakine level to stimulate the hemocytes proliferation. Altogether, we identified the shrimp crustin Pm4 and STG I as novel RNA binding proteins that play an important role in down-regulating astakine expression at post-transcriptional level and are crucial for the maintenance of hematopoiesis.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Proteínas de Artrópodes/metabolismo , Decápodes/metabolismo , Regulação para Baixo/fisiologia , Hematopoese/fisiologia , Ribonucleoproteínas/metabolismo , Transglutaminases/metabolismo , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/biossíntese , Regiões 3' não Traduzidas/fisiologia , Animais , Peptídeos Catiônicos Antimicrobianos/genética , Proteínas de Artrópodes/genética , Decápodes/genética , Hemócitos/citologia , Hemócitos/metabolismo , Estabilidade de RNA/fisiologia , Ribonucleoproteínas/genética , Células Sf9 , Spodoptera , Transglutaminases/genética , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/genética
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