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Actin, RhoA, and Rab11 participation during encystment in Entamoeba invadens.

Herrera-Martínez M, Hernández-Ramírez VI, Lagunes-Guillén AE, Chávez-Munguía B, Talamás-Rohana P - Biomed Res Int (2013)

Bottom Line: Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles.Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%.These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del I.P.N., Avenida Instituto Politécnico Nacional No. 2508, Colonia San Pedro Zacatenco, Delegación Gustavo A. Madero, 07360 México City, DF, Mexico.

ABSTRACT
In the genus Entamoeba, actin reorganization is necessary for cyst differentiation; however, its role is still unknown. The aim of this work was to investigate the role of actin and encystation-related proteins during Entamoeba invadens encystation. Studied proteins were actin, RhoA, a small GTPase involved through its effectors in the rearrangement of the actin cytoskeleton; Rab11, a protein involved in the transport of encystation vesicles; and enolase, as an encystment vesicles marker. Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles. Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%. These inhibitors also blocked cell rounding, disposal of vacuoles, and the proper formation of the cysts wall. At later times, F-actin and Rab11 colocalized with enolase, suggesting that Rab11 could participate in the transport of the cyst wall components through the F-actin cytoskeleton. These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens.

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Related in: MedlinePlus

Bioinformatic analysis of sequence homology between Eiactin, EiRab11, and Eienolase with their human counterparts molecules. (a) High identity of N-terminal between Eiactin, Hsactin, and Ggactin is shown. (b) EiRab11 presented an identity of 54–57% with HsRab11A on their complete sequence. (c) Eienolase presented high identity of its N-terminal sequence with human enolase residues. Identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.”.
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fig1: Bioinformatic analysis of sequence homology between Eiactin, EiRab11, and Eienolase with their human counterparts molecules. (a) High identity of N-terminal between Eiactin, Hsactin, and Ggactin is shown. (b) EiRab11 presented an identity of 54–57% with HsRab11A on their complete sequence. (c) Eienolase presented high identity of its N-terminal sequence with human enolase residues. Identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.”.

Mentions: As a first approach for this study, bioinformatic analysis of the molecules of Entamoeba invadens and Homo sapiens was done to ensure that heterologous antibodies could be used to track the proteins of interest. Clustal 2.1 multiple sequence alignment program was used; identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.” (Figure 1). Eiactin presented 85–88% identity with Hsactin and Ggactin (Gallus gallus) based on their complete sequences. High identity of N-terminal between all molecules is shown in Figure 1(a). EiRab11 presented a 54–57% identity with HsRab11A based on their complete sequences. Identity of complete sequences is shown (Figure 1(b)). Eienolase presented 64% identity with Hsalphaenolase on the basis of the complete sequence. High identity of N-terminal between all molecules is shown (Figure 1(c)). Blast analysis (PubMed) of RhoA of human origin was run to find similar sequences in E. invadens. First 10 sequences were taken to perform an alignment of them. EiRho presented 47–51% identity with HsRhoA. Based on the above results, it was possible to predict a substantial cross-reactivity between anti-human proteins antibodies and homologous isoforms in E. invadens.


Actin, RhoA, and Rab11 participation during encystment in Entamoeba invadens.

Herrera-Martínez M, Hernández-Ramírez VI, Lagunes-Guillén AE, Chávez-Munguía B, Talamás-Rohana P - Biomed Res Int (2013)

Bioinformatic analysis of sequence homology between Eiactin, EiRab11, and Eienolase with their human counterparts molecules. (a) High identity of N-terminal between Eiactin, Hsactin, and Ggactin is shown. (b) EiRab11 presented an identity of 54–57% with HsRab11A on their complete sequence. (c) Eienolase presented high identity of its N-terminal sequence with human enolase residues. Identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.”.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3794519&req=5

fig1: Bioinformatic analysis of sequence homology between Eiactin, EiRab11, and Eienolase with their human counterparts molecules. (a) High identity of N-terminal between Eiactin, Hsactin, and Ggactin is shown. (b) EiRab11 presented an identity of 54–57% with HsRab11A on their complete sequence. (c) Eienolase presented high identity of its N-terminal sequence with human enolase residues. Identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.”.
Mentions: As a first approach for this study, bioinformatic analysis of the molecules of Entamoeba invadens and Homo sapiens was done to ensure that heterologous antibodies could be used to track the proteins of interest. Clustal 2.1 multiple sequence alignment program was used; identical residues are shown in “∗”, conserved residues in “:”, and semiconserved residues in “.” (Figure 1). Eiactin presented 85–88% identity with Hsactin and Ggactin (Gallus gallus) based on their complete sequences. High identity of N-terminal between all molecules is shown in Figure 1(a). EiRab11 presented a 54–57% identity with HsRab11A based on their complete sequences. Identity of complete sequences is shown (Figure 1(b)). Eienolase presented 64% identity with Hsalphaenolase on the basis of the complete sequence. High identity of N-terminal between all molecules is shown (Figure 1(c)). Blast analysis (PubMed) of RhoA of human origin was run to find similar sequences in E. invadens. First 10 sequences were taken to perform an alignment of them. EiRho presented 47–51% identity with HsRhoA. Based on the above results, it was possible to predict a substantial cross-reactivity between anti-human proteins antibodies and homologous isoforms in E. invadens.

Bottom Line: Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles.Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%.These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del I.P.N., Avenida Instituto Politécnico Nacional No. 2508, Colonia San Pedro Zacatenco, Delegación Gustavo A. Madero, 07360 México City, DF, Mexico.

ABSTRACT
In the genus Entamoeba, actin reorganization is necessary for cyst differentiation; however, its role is still unknown. The aim of this work was to investigate the role of actin and encystation-related proteins during Entamoeba invadens encystation. Studied proteins were actin, RhoA, a small GTPase involved through its effectors in the rearrangement of the actin cytoskeleton; Rab11, a protein involved in the transport of encystation vesicles; and enolase, as an encystment vesicles marker. Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles. Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%. These inhibitors also blocked cell rounding, disposal of vacuoles, and the proper formation of the cysts wall. At later times, F-actin and Rab11 colocalized with enolase, suggesting that Rab11 could participate in the transport of the cyst wall components through the F-actin cytoskeleton. These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens.

Show MeSH
Related in: MedlinePlus