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Identification and expression analysis of MATE genes involved in flavonoid transport in blueberry plants.

Chen L, Liu Y, Liu H, Kang L, Geng J, Gai Y, Ding Y, Sun H, Li Y - PLoS ONE (2015)

Bottom Line: Here 33 Unigenes annotated as MATE transporters were found in the blueberry fruit transcriptome, of which eight full-length cDNA sequences were identified and cloned.These results showed VcMATEs might be involved in diverse physiological functions, and anthocyanins across the membranes might be mutually maintained by MATE-type flavonoid transporters and other mechanisms.This study will enrich the MATE-based transport mechanisms of secondary metabolite, and provide a new biotechonology strategy to develop better nutritional blueberry cultivars.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Jilin Agricultural University, Changchun, Jilin, China.

ABSTRACT
Multidrug and toxic compound extrusion (MATE) proteins are the most recently identified family of multidrug transporters. In plants, this family is remarkably large compared to the human and bacteria counterpart, highlighting the importance of MATE proteins in this kingdom. Here 33 Unigenes annotated as MATE transporters were found in the blueberry fruit transcriptome, of which eight full-length cDNA sequences were identified and cloned. These proteins are composed of 477-517 residues, with molecular masses ~54 kDa, and theoretical isoelectric points from 5.35 to 8.41. Bioinformatics analysis predicted 10-12 putative transmembrane segments for VcMATEs, and localization to the plasma membrane without an N-terminal signal peptide. All blueberry MATE proteins shared 32.1-84.4% identity, among which VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8, and VcMATE9 were more similar to the MATE-type flavonoid transporters. Phylogenetic analysis showed VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8 and VcMATE9 clustered with MATE-type flavonoid transporters, indicating that they might be involved in flavonoid transport. VcMATE1 and VcMATE4 may be involved in the transport of secondary metabolites, the detoxification of xenobiotics, or the export of toxic cations. Real-time quantitative PCR demonstrated that the expression profile of the eight VcMATE genes varied spatially and temporally. Analysis of expression and anthocyanin accumulation indicated that there were some correlation between the expression profile and the accumulation of anthocyanins. These results showed VcMATEs might be involved in diverse physiological functions, and anthocyanins across the membranes might be mutually maintained by MATE-type flavonoid transporters and other mechanisms. This study will enrich the MATE-based transport mechanisms of secondary metabolite, and provide a new biotechonology strategy to develop better nutritional blueberry cultivars.

No MeSH data available.


Secondary structures of the 8 blueberry MATE proteins predicted by SOPMA.The α-helix, extended strand, β-turn, and random coil are denoted as the longest (blue), medium long (dark red), short (green), and the shortest (pink) vertical bars, respectively.
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pone.0118578.g001: Secondary structures of the 8 blueberry MATE proteins predicted by SOPMA.The α-helix, extended strand, β-turn, and random coil are denoted as the longest (blue), medium long (dark red), short (green), and the shortest (pink) vertical bars, respectively.

Mentions: As presented in Table 3, VcMATE cDNA encoded proteins are composed of 477–537 residues, with predicted molecular masses around 54 kDa, theoretical isoelectric points from 4.89 to 8.41, aliphatic indices between 113.88 and 126.29, and grand average of hydropathicity around 0.7. All deduced VcMATE proteins were predicted as stable because their instability index is smaller than 40. When analyzed by SOPMA, the secondary structures of blueberry MATEs were predicted to be mainly composed of alpha helices (60.34–68.13%) interspersed with random coils (18.13–22.35%), extended strands (10.36–14.53%), and beta turns (2.39–4.09%) (Fig. 1). For each protein, there were more than 15 distinct alpha helices dispersed along the length of the protein. The prediction of transmembrane domains using HMMTOP suggested that there were 10–12 putative transmembrane (TM) segments in VcMATEs (Fig. 2). Prediction of the subcellular localization of VcMATEs was performed using the PSORT program. The results showed that the eight transporters were all localized to the plasma membrane (Table 3). Proteins are usually guided to their target membranes by signal peptides, but there were no signal peptides in the N-terminus according to SignalP. It is possible that signal peptides do not have strictly conserved amino acid sequences and are thus sometimes difficult to identify.


Identification and expression analysis of MATE genes involved in flavonoid transport in blueberry plants.

Chen L, Liu Y, Liu H, Kang L, Geng J, Gai Y, Ding Y, Sun H, Li Y - PLoS ONE (2015)

Secondary structures of the 8 blueberry MATE proteins predicted by SOPMA.The α-helix, extended strand, β-turn, and random coil are denoted as the longest (blue), medium long (dark red), short (green), and the shortest (pink) vertical bars, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363705&req=5

pone.0118578.g001: Secondary structures of the 8 blueberry MATE proteins predicted by SOPMA.The α-helix, extended strand, β-turn, and random coil are denoted as the longest (blue), medium long (dark red), short (green), and the shortest (pink) vertical bars, respectively.
Mentions: As presented in Table 3, VcMATE cDNA encoded proteins are composed of 477–537 residues, with predicted molecular masses around 54 kDa, theoretical isoelectric points from 4.89 to 8.41, aliphatic indices between 113.88 and 126.29, and grand average of hydropathicity around 0.7. All deduced VcMATE proteins were predicted as stable because their instability index is smaller than 40. When analyzed by SOPMA, the secondary structures of blueberry MATEs were predicted to be mainly composed of alpha helices (60.34–68.13%) interspersed with random coils (18.13–22.35%), extended strands (10.36–14.53%), and beta turns (2.39–4.09%) (Fig. 1). For each protein, there were more than 15 distinct alpha helices dispersed along the length of the protein. The prediction of transmembrane domains using HMMTOP suggested that there were 10–12 putative transmembrane (TM) segments in VcMATEs (Fig. 2). Prediction of the subcellular localization of VcMATEs was performed using the PSORT program. The results showed that the eight transporters were all localized to the plasma membrane (Table 3). Proteins are usually guided to their target membranes by signal peptides, but there were no signal peptides in the N-terminus according to SignalP. It is possible that signal peptides do not have strictly conserved amino acid sequences and are thus sometimes difficult to identify.

Bottom Line: Here 33 Unigenes annotated as MATE transporters were found in the blueberry fruit transcriptome, of which eight full-length cDNA sequences were identified and cloned.These results showed VcMATEs might be involved in diverse physiological functions, and anthocyanins across the membranes might be mutually maintained by MATE-type flavonoid transporters and other mechanisms.This study will enrich the MATE-based transport mechanisms of secondary metabolite, and provide a new biotechonology strategy to develop better nutritional blueberry cultivars.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Jilin Agricultural University, Changchun, Jilin, China.

ABSTRACT
Multidrug and toxic compound extrusion (MATE) proteins are the most recently identified family of multidrug transporters. In plants, this family is remarkably large compared to the human and bacteria counterpart, highlighting the importance of MATE proteins in this kingdom. Here 33 Unigenes annotated as MATE transporters were found in the blueberry fruit transcriptome, of which eight full-length cDNA sequences were identified and cloned. These proteins are composed of 477-517 residues, with molecular masses ~54 kDa, and theoretical isoelectric points from 5.35 to 8.41. Bioinformatics analysis predicted 10-12 putative transmembrane segments for VcMATEs, and localization to the plasma membrane without an N-terminal signal peptide. All blueberry MATE proteins shared 32.1-84.4% identity, among which VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8, and VcMATE9 were more similar to the MATE-type flavonoid transporters. Phylogenetic analysis showed VcMATE2, VcMATE3, VcMATE5, VcMATE7, VcMATE8 and VcMATE9 clustered with MATE-type flavonoid transporters, indicating that they might be involved in flavonoid transport. VcMATE1 and VcMATE4 may be involved in the transport of secondary metabolites, the detoxification of xenobiotics, or the export of toxic cations. Real-time quantitative PCR demonstrated that the expression profile of the eight VcMATE genes varied spatially and temporally. Analysis of expression and anthocyanin accumulation indicated that there were some correlation between the expression profile and the accumulation of anthocyanins. These results showed VcMATEs might be involved in diverse physiological functions, and anthocyanins across the membranes might be mutually maintained by MATE-type flavonoid transporters and other mechanisms. This study will enrich the MATE-based transport mechanisms of secondary metabolite, and provide a new biotechonology strategy to develop better nutritional blueberry cultivars.

No MeSH data available.