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Multicopper oxidase-1 is required for iron homeostasis in Malpighian tubules of Helicoverpa armigera.

Liu X, Sun C, Liu X, Yin X, Wang B, Du M, An S - Sci Rep (2015)

Bottom Line: HaMCO1 was also found to be highly abundant in Malpighian tubules.HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression.Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules.

View Article: PubMed Central - PubMed

Affiliation: State key Laboratory of Wheat and Maize Crop Science/College of Plant Protection, Henan Agricultural University, Zhengzhou 450002 P.R. China.

ABSTRACT
Multicopper oxidases (MCOs) are enzymes that contain 10 conserved histidine residues and 1 cysteine residue. MCO1 has been extensively investigated in the midgut because this MCO is implicated in ascorbate oxidation, iron homeostasis and immune responses. However, information regarding the action of MCO1 in Malpighian tubules is limited. In this study, Helicoverpa armigera was used as a model to investigate the function of MCO1 in Malpighian tubules. Sequence analysis results revealed that HaMCO1 exhibits typical MCO characteristics, with 10 histidine and 1 cysteine residues for copper ion binding. HaMCO1 was also found to be highly abundant in Malpighian tubules. Temporal expression patterns indicated that HaMCO1 is mainly expressed during larval molting stages. Hormone treatments [the molting hormone 20-hydroxyecdysone (20E) and juvenile hormone (JH)] revealed that 20E inhibits HaMCO1 transcript expression via its heterodimer receptor, which consists of ecdysone receptor (EcR) and ultraspiracle (USP), and that JH counteracts the action of 20E to activate HaMCO1 transcript expression via its intracellular receptor methoprene-tolerant (Met). HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression. Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules.

No MeSH data available.


Nucleotide sequence and putative amino acid sequence of HaMCO1.The underlining indicates the initiation codon or stop codon. The predicted signal peptide is indicated in italicized text. The putative carboxyl-terminal transmembrane region is delineated with a blue line.
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f1: Nucleotide sequence and putative amino acid sequence of HaMCO1.The underlining indicates the initiation codon or stop codon. The predicted signal peptide is indicated in italicized text. The putative carboxyl-terminal transmembrane region is delineated with a blue line.

Mentions: The MCO1 sequence designated HaMCO1 was obtained from transcriptome data of H. armigera (data not shown). HaMCO1 contains an open reading frame (ORF) of 2,430 bp, which encodes a putative protein of 810 amino acid residues, with a molecular weight with 91.998 kDa and an isoelectric point of 5.46 (Fig. 1). Similar to MCO1 in other insects, HaCOM1 consists of a secretion signal peptide sequence (located at amino acids 1–23 of the HaMCO1 primary sequence), and a carboxyl-terminal transmembrane region (located at amino acids 794–809 of HaMCO1) (Fig. 1); HaMCO1 is predicted to be GPI-anchored. Most importantly, HaMCO1 also contains ten histidines and one cysteine, which are typical characteristics of MCOs; these residues are required for copper ion binding (Fig. 2). The HaMCO1 amino acid sequence was subjected to further multiple sequence alignments with homologous proteins. The results also revealed that HaMCO1 contains ten histidine residues and one cysteine residue (Fig. 2). Therefore, the sequence obtained from the transcriptome data corresponds to an MCO. Homology analysis showed that HaMCO1 shares 84% amino acid sequence identity with Danaus plexippus laccase1, 79% amino acid sequence identity with Bombyx mori MCO1, and 75% amino acid sequence identity with M. sexta MCO1. Indeed, our obtained HaMCO encodes an MCO1. Phylogenetic analysis also revealed that HaMCO1 clusters with the MCO1 of other insects (Fig. 3); hence, the obtained HaMCO1 sequence is reliable.


Multicopper oxidase-1 is required for iron homeostasis in Malpighian tubules of Helicoverpa armigera.

Liu X, Sun C, Liu X, Yin X, Wang B, Du M, An S - Sci Rep (2015)

Nucleotide sequence and putative amino acid sequence of HaMCO1.The underlining indicates the initiation codon or stop codon. The predicted signal peptide is indicated in italicized text. The putative carboxyl-terminal transmembrane region is delineated with a blue line.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4593997&req=5

f1: Nucleotide sequence and putative amino acid sequence of HaMCO1.The underlining indicates the initiation codon or stop codon. The predicted signal peptide is indicated in italicized text. The putative carboxyl-terminal transmembrane region is delineated with a blue line.
Mentions: The MCO1 sequence designated HaMCO1 was obtained from transcriptome data of H. armigera (data not shown). HaMCO1 contains an open reading frame (ORF) of 2,430 bp, which encodes a putative protein of 810 amino acid residues, with a molecular weight with 91.998 kDa and an isoelectric point of 5.46 (Fig. 1). Similar to MCO1 in other insects, HaCOM1 consists of a secretion signal peptide sequence (located at amino acids 1–23 of the HaMCO1 primary sequence), and a carboxyl-terminal transmembrane region (located at amino acids 794–809 of HaMCO1) (Fig. 1); HaMCO1 is predicted to be GPI-anchored. Most importantly, HaMCO1 also contains ten histidines and one cysteine, which are typical characteristics of MCOs; these residues are required for copper ion binding (Fig. 2). The HaMCO1 amino acid sequence was subjected to further multiple sequence alignments with homologous proteins. The results also revealed that HaMCO1 contains ten histidine residues and one cysteine residue (Fig. 2). Therefore, the sequence obtained from the transcriptome data corresponds to an MCO. Homology analysis showed that HaMCO1 shares 84% amino acid sequence identity with Danaus plexippus laccase1, 79% amino acid sequence identity with Bombyx mori MCO1, and 75% amino acid sequence identity with M. sexta MCO1. Indeed, our obtained HaMCO encodes an MCO1. Phylogenetic analysis also revealed that HaMCO1 clusters with the MCO1 of other insects (Fig. 3); hence, the obtained HaMCO1 sequence is reliable.

Bottom Line: HaMCO1 was also found to be highly abundant in Malpighian tubules.HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression.Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules.

View Article: PubMed Central - PubMed

Affiliation: State key Laboratory of Wheat and Maize Crop Science/College of Plant Protection, Henan Agricultural University, Zhengzhou 450002 P.R. China.

ABSTRACT
Multicopper oxidases (MCOs) are enzymes that contain 10 conserved histidine residues and 1 cysteine residue. MCO1 has been extensively investigated in the midgut because this MCO is implicated in ascorbate oxidation, iron homeostasis and immune responses. However, information regarding the action of MCO1 in Malpighian tubules is limited. In this study, Helicoverpa armigera was used as a model to investigate the function of MCO1 in Malpighian tubules. Sequence analysis results revealed that HaMCO1 exhibits typical MCO characteristics, with 10 histidine and 1 cysteine residues for copper ion binding. HaMCO1 was also found to be highly abundant in Malpighian tubules. Temporal expression patterns indicated that HaMCO1 is mainly expressed during larval molting stages. Hormone treatments [the molting hormone 20-hydroxyecdysone (20E) and juvenile hormone (JH)] revealed that 20E inhibits HaMCO1 transcript expression via its heterodimer receptor, which consists of ecdysone receptor (EcR) and ultraspiracle (USP), and that JH counteracts the action of 20E to activate HaMCO1 transcript expression via its intracellular receptor methoprene-tolerant (Met). HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression. Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules.

No MeSH data available.