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Identification and comparative analysis of subolesin/akirin ortholog from Ornithodoros turicata ticks.

Sultana H, Patel U, Sonenshine DE, Neelakanta G - Parasit Vectors (2015)

Bottom Line: Our study reveals that O. turicata subolesin gene expression is developmentally regulated, where; adult ticks expressed significantly higher levels in comparison to the larvae or nymphal ticks.However, no myristoylation or glycosylation sites were evident in the O. turicata subolesin sequence.Our study provides important insights in recognizing subolesin as a conserved potential candidate for the development of a broad-spectrum anti-vector vaccine to control not only ticks but also several other arthropods that transmit diseases to humans and animals.

View Article: PubMed Central - PubMed

Affiliation: Center for Molecular Medicine, College of Sciences, Old Dominion University, Norfolk, 23529, VA, USA. hsultana@odu.edu.

ABSTRACT

Background: Subolesin is an evolutionary conserved molecule in diverse arthropod species that play an important role in the regulation of genes involved in immune responses, blood digestion, reproduction and development. In this study, we have identified a subolesin ortholog from soft ticks Ornithodoros turicata, the vector of the relapsing fever spirochete in the United States.

Methods: Uninfected fed or unfed O. turicata ticks were used throughout this study. The subolesin mRNA was amplified by reverse transcription polymerase chain reaction (RT-PCR) and sequenced. Quantitative-real time PCR (QRT-PCR) was performed to evaluate subolesin mRNA levels at different O. turicata developmental stages and from salivary glands and gut tissues. Bioinformatics and comparative analysis was performed to predict potential post-translational modifications in O. turicata subolesin amino-acid sequences.

Results: Our study reveals that O. turicata subolesin gene expression is developmentally regulated, where; adult ticks expressed significantly higher levels in comparison to the larvae or nymphal ticks. Expression of subolesin was evident in both unfed and fed ticks and in the salivary glands and midgut tissues. The expression of subolesin transcripts varied in fed ticks with peak levels at day 14 post-feeding. Phylogenetic analysis revealed that O. turicata subolesin showed a high degree of sequence conservation with subolesin's from other soft and hard ticks. Bioinformatics and comparative analysis predicted that O. turicata subolesin carry three Protein kinase C and one Casein kinase II phosphorylation sites. However, no myristoylation or glycosylation sites were evident in the O. turicata subolesin sequence.

Conclusion: Our study provides important insights in recognizing subolesin as a conserved potential candidate for the development of a broad-spectrum anti-vector vaccine to control not only ticks but also several other arthropods that transmit diseases to humans and animals.

No MeSH data available.


Related in: MedlinePlus

Prediction and comparative analysis ofO. turicatasubolesin post-translational modifications. Annotated amino acid sequences of several subolesins were individually analyzed in PROSITE database for PKC phosphorylation (A), CK2 Phosphorylation (B), Myristoylation (C) and Glycosylation (D) sites. Histograms represent number of post-translational modification sites for each Subolesin. Organism names and groups are shown at the bottom of the figure.
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Fig5: Prediction and comparative analysis ofO. turicatasubolesin post-translational modifications. Annotated amino acid sequences of several subolesins were individually analyzed in PROSITE database for PKC phosphorylation (A), CK2 Phosphorylation (B), Myristoylation (C) and Glycosylation (D) sites. Histograms represent number of post-translational modification sites for each Subolesin. Organism names and groups are shown at the bottom of the figure.

Mentions: To predict conserved and/or unique post-translational modifications, subolesin amino acid sequences from hard and soft ticks, mosquitos and Glossina species were analyzed by PROSITE as described in the Methods section. Based on the PROSITE analysis, all subolesin sequences carried more than two Protein kinase C (PKC) phosphorylation sites (Figure 5A). C. quinquefasciatus subolesin sequence showed higher numbers (seven) and H. longicornis and O. turicata ticks showed lower numbers (three) of PKC phosphorylation sites (Figure 5A). In addition, all subolesin sequences carried at least one Casein kinase II (CK2) phosphorylation site (Figure 5B). The O. erraticus soft ticks carried higher numbers (three) and three mosquito species (A. aegypti, A. albopictus, A. gambiae) and O. turicata ticks carried one CK2 phosphorylation site (Figure 5B). The number of myristoylation sites in the subolesin sequences varied among different species (Figure 5C). One hard tick (H. marginatum) and two soft ticks (O. erraticus and O. turicata) had no myristoylation sites in the sequence (Figure 5C). Whereas, hard tick H. qinghaiensis carried large number of myristoylation sites (five) in comparison to all other arthropod species (Figure 5C). Prediction of glycosylation sites in different subolesins revealed that, except A. aegypti all other mosquito subolesins that were analyzed carried at least one glycosylation site in the sequence (Figure 5D). Among hard and soft ticks, O. erraticus is the only tick that carried glycosylation site in the subolesin sequence (Figure 5D). Among all the members, the A. aegypti and A. gambiae subolesin’s are the only two that carry serine-rich domain at N-terminus of the sequence (Additional file 1: Figure S2). Collectively, predictions of the post-translational modifications revealed interesting insights for the molecular function of subolesins/akirins in various arthropod species that transmit human pathogens.Figure 5


Identification and comparative analysis of subolesin/akirin ortholog from Ornithodoros turicata ticks.

Sultana H, Patel U, Sonenshine DE, Neelakanta G - Parasit Vectors (2015)

Prediction and comparative analysis ofO. turicatasubolesin post-translational modifications. Annotated amino acid sequences of several subolesins were individually analyzed in PROSITE database for PKC phosphorylation (A), CK2 Phosphorylation (B), Myristoylation (C) and Glycosylation (D) sites. Histograms represent number of post-translational modification sites for each Subolesin. Organism names and groups are shown at the bottom of the figure.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359563&req=5

Fig5: Prediction and comparative analysis ofO. turicatasubolesin post-translational modifications. Annotated amino acid sequences of several subolesins were individually analyzed in PROSITE database for PKC phosphorylation (A), CK2 Phosphorylation (B), Myristoylation (C) and Glycosylation (D) sites. Histograms represent number of post-translational modification sites for each Subolesin. Organism names and groups are shown at the bottom of the figure.
Mentions: To predict conserved and/or unique post-translational modifications, subolesin amino acid sequences from hard and soft ticks, mosquitos and Glossina species were analyzed by PROSITE as described in the Methods section. Based on the PROSITE analysis, all subolesin sequences carried more than two Protein kinase C (PKC) phosphorylation sites (Figure 5A). C. quinquefasciatus subolesin sequence showed higher numbers (seven) and H. longicornis and O. turicata ticks showed lower numbers (three) of PKC phosphorylation sites (Figure 5A). In addition, all subolesin sequences carried at least one Casein kinase II (CK2) phosphorylation site (Figure 5B). The O. erraticus soft ticks carried higher numbers (three) and three mosquito species (A. aegypti, A. albopictus, A. gambiae) and O. turicata ticks carried one CK2 phosphorylation site (Figure 5B). The number of myristoylation sites in the subolesin sequences varied among different species (Figure 5C). One hard tick (H. marginatum) and two soft ticks (O. erraticus and O. turicata) had no myristoylation sites in the sequence (Figure 5C). Whereas, hard tick H. qinghaiensis carried large number of myristoylation sites (five) in comparison to all other arthropod species (Figure 5C). Prediction of glycosylation sites in different subolesins revealed that, except A. aegypti all other mosquito subolesins that were analyzed carried at least one glycosylation site in the sequence (Figure 5D). Among hard and soft ticks, O. erraticus is the only tick that carried glycosylation site in the subolesin sequence (Figure 5D). Among all the members, the A. aegypti and A. gambiae subolesin’s are the only two that carry serine-rich domain at N-terminus of the sequence (Additional file 1: Figure S2). Collectively, predictions of the post-translational modifications revealed interesting insights for the molecular function of subolesins/akirins in various arthropod species that transmit human pathogens.Figure 5

Bottom Line: Our study reveals that O. turicata subolesin gene expression is developmentally regulated, where; adult ticks expressed significantly higher levels in comparison to the larvae or nymphal ticks.However, no myristoylation or glycosylation sites were evident in the O. turicata subolesin sequence.Our study provides important insights in recognizing subolesin as a conserved potential candidate for the development of a broad-spectrum anti-vector vaccine to control not only ticks but also several other arthropods that transmit diseases to humans and animals.

View Article: PubMed Central - PubMed

Affiliation: Center for Molecular Medicine, College of Sciences, Old Dominion University, Norfolk, 23529, VA, USA. hsultana@odu.edu.

ABSTRACT

Background: Subolesin is an evolutionary conserved molecule in diverse arthropod species that play an important role in the regulation of genes involved in immune responses, blood digestion, reproduction and development. In this study, we have identified a subolesin ortholog from soft ticks Ornithodoros turicata, the vector of the relapsing fever spirochete in the United States.

Methods: Uninfected fed or unfed O. turicata ticks were used throughout this study. The subolesin mRNA was amplified by reverse transcription polymerase chain reaction (RT-PCR) and sequenced. Quantitative-real time PCR (QRT-PCR) was performed to evaluate subolesin mRNA levels at different O. turicata developmental stages and from salivary glands and gut tissues. Bioinformatics and comparative analysis was performed to predict potential post-translational modifications in O. turicata subolesin amino-acid sequences.

Results: Our study reveals that O. turicata subolesin gene expression is developmentally regulated, where; adult ticks expressed significantly higher levels in comparison to the larvae or nymphal ticks. Expression of subolesin was evident in both unfed and fed ticks and in the salivary glands and midgut tissues. The expression of subolesin transcripts varied in fed ticks with peak levels at day 14 post-feeding. Phylogenetic analysis revealed that O. turicata subolesin showed a high degree of sequence conservation with subolesin's from other soft and hard ticks. Bioinformatics and comparative analysis predicted that O. turicata subolesin carry three Protein kinase C and one Casein kinase II phosphorylation sites. However, no myristoylation or glycosylation sites were evident in the O. turicata subolesin sequence.

Conclusion: Our study provides important insights in recognizing subolesin as a conserved potential candidate for the development of a broad-spectrum anti-vector vaccine to control not only ticks but also several other arthropods that transmit diseases to humans and animals.

No MeSH data available.


Related in: MedlinePlus