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ANKS3 Co-Localises with ANKS6 in Mouse Renal Cilia and Is Associated with Vasopressin Signaling and Apoptosis In Vivo in Mice.

Delestré L, Bakey Z, Prado C, Hoffmann S, Bihoreau MT, Lelongt B, Gauguier D - PLoS ONE (2015)

Bottom Line: Following experimental confirmation of interaction between ANKS6and ANKS3 using a Yeast two hybrid system, we demonstrated that binding between the two proteins occurs through their sterile alpha motif (SAM) and that the amino acid 823 in rat ANSK6 is key for this interaction.Downregulated expression of Anks3 in vivo in mice by Locked Nucleic Acid (LNA) modified antisense oligonucleotides was associated with increased transcription of vasopressin-induced genes, suggesting changes in renal water permeability, and altered transcription of genes encoding proteins involved in cilium structure, apoptosis and cell proliferation.Our results contribute to improved knowledge of the structure and function of the network of proteins interacting with ANKS6, which may represent therapeutic targets in cystic diseases.

View Article: PubMed Central - PubMed

Affiliation: Sorbonne Universities, University Pierre and Marie Curie, University Paris Descartes, Sorbonne Paris Cité, INSERM, UMR_S1138, Cordeliers Research Centre, Paris, France.

ABSTRACT
Mutations in Ankyrin repeat and sterile alpha motif domain containing 6 (ANKS6) play a causative role in renal cyst formation in the PKD/Mhm(cy/+) rat model of polycystic kidney disease and in nephronophthisis in humans. A network of protein partners of ANKS6 is emerging and their functional characterization provides important clues to understand the role of ANKS6 in renal biology and in mechanisms involved in the formation of renal cysts. Following experimental confirmation of interaction between ANKS6and ANKS3 using a Yeast two hybrid system, we demonstrated that binding between the two proteins occurs through their sterile alpha motif (SAM) and that the amino acid 823 in rat ANSK6 is key for this interaction. We further showed their interaction by co-immunoprecipitation and showed in vivo in mice that ANKS3 is present in renal cilia. Downregulated expression of Anks3 in vivo in mice by Locked Nucleic Acid (LNA) modified antisense oligonucleotides was associated with increased transcription of vasopressin-induced genes, suggesting changes in renal water permeability, and altered transcription of genes encoding proteins involved in cilium structure, apoptosis and cell proliferation. These data provide experimental evidence of ANKS3-ANKS6 direct interaction through their SAM domain and co-localisation in mouse renal cilia, and shed light on molecular mechanisms indirectly mediated by ANKS6 in the mouse kidney, that may be affected by altered ANKS3-ANKS6 interaction. Our results contribute to improved knowledge of the structure and function of the network of proteins interacting with ANKS6, which may represent therapeutic targets in cystic diseases.

No MeSH data available.


Related in: MedlinePlus

The mutation (R823W) in PKD/Mhm(cy/cy) rats does not alter ANKS6-ANKS3 renal co-localisation.Immunohistochemical staining of renal cortex and medulla was performed in 3 week-old wild-type and PKD/Mhm(cy/cy) rats for ANKS3 (upper panel) and ANKS6 (lower panel).
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pone.0136781.g004: The mutation (R823W) in PKD/Mhm(cy/cy) rats does not alter ANKS6-ANKS3 renal co-localisation.Immunohistochemical staining of renal cortex and medulla was performed in 3 week-old wild-type and PKD/Mhm(cy/cy) rats for ANKS3 (upper panel) and ANKS6 (lower panel).

Mentions: Subsequent immunohistological studies in mouse kidneys using specific markers of nephron segments revealed co-localisation of ANKS6 and ANKS3 in glomeruli and in tubules (Fig 3C). Direct interaction between the two proteins in the kidney was confirmed by co-immunoprecipitation of ANKS3 and ANKS6 on kidney lysates with immobilized anti-ANKS3 or anti-ANKS6 antibody (Fig 3D). Finally, we analysed the effect of the R823W mutation in ANKS6 on renal localisation of ANKS3 and ANKS6 and showed that the two proteins remain present in collecting duct cells of renal medulla in both wild-type and cy/cy rats (Fig 4).


ANKS3 Co-Localises with ANKS6 in Mouse Renal Cilia and Is Associated with Vasopressin Signaling and Apoptosis In Vivo in Mice.

Delestré L, Bakey Z, Prado C, Hoffmann S, Bihoreau MT, Lelongt B, Gauguier D - PLoS ONE (2015)

The mutation (R823W) in PKD/Mhm(cy/cy) rats does not alter ANKS6-ANKS3 renal co-localisation.Immunohistochemical staining of renal cortex and medulla was performed in 3 week-old wild-type and PKD/Mhm(cy/cy) rats for ANKS3 (upper panel) and ANKS6 (lower panel).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556665&req=5

pone.0136781.g004: The mutation (R823W) in PKD/Mhm(cy/cy) rats does not alter ANKS6-ANKS3 renal co-localisation.Immunohistochemical staining of renal cortex and medulla was performed in 3 week-old wild-type and PKD/Mhm(cy/cy) rats for ANKS3 (upper panel) and ANKS6 (lower panel).
Mentions: Subsequent immunohistological studies in mouse kidneys using specific markers of nephron segments revealed co-localisation of ANKS6 and ANKS3 in glomeruli and in tubules (Fig 3C). Direct interaction between the two proteins in the kidney was confirmed by co-immunoprecipitation of ANKS3 and ANKS6 on kidney lysates with immobilized anti-ANKS3 or anti-ANKS6 antibody (Fig 3D). Finally, we analysed the effect of the R823W mutation in ANKS6 on renal localisation of ANKS3 and ANKS6 and showed that the two proteins remain present in collecting duct cells of renal medulla in both wild-type and cy/cy rats (Fig 4).

Bottom Line: Following experimental confirmation of interaction between ANKS6and ANKS3 using a Yeast two hybrid system, we demonstrated that binding between the two proteins occurs through their sterile alpha motif (SAM) and that the amino acid 823 in rat ANSK6 is key for this interaction.Downregulated expression of Anks3 in vivo in mice by Locked Nucleic Acid (LNA) modified antisense oligonucleotides was associated with increased transcription of vasopressin-induced genes, suggesting changes in renal water permeability, and altered transcription of genes encoding proteins involved in cilium structure, apoptosis and cell proliferation.Our results contribute to improved knowledge of the structure and function of the network of proteins interacting with ANKS6, which may represent therapeutic targets in cystic diseases.

View Article: PubMed Central - PubMed

Affiliation: Sorbonne Universities, University Pierre and Marie Curie, University Paris Descartes, Sorbonne Paris Cité, INSERM, UMR_S1138, Cordeliers Research Centre, Paris, France.

ABSTRACT
Mutations in Ankyrin repeat and sterile alpha motif domain containing 6 (ANKS6) play a causative role in renal cyst formation in the PKD/Mhm(cy/+) rat model of polycystic kidney disease and in nephronophthisis in humans. A network of protein partners of ANKS6 is emerging and their functional characterization provides important clues to understand the role of ANKS6 in renal biology and in mechanisms involved in the formation of renal cysts. Following experimental confirmation of interaction between ANKS6and ANKS3 using a Yeast two hybrid system, we demonstrated that binding between the two proteins occurs through their sterile alpha motif (SAM) and that the amino acid 823 in rat ANSK6 is key for this interaction. We further showed their interaction by co-immunoprecipitation and showed in vivo in mice that ANKS3 is present in renal cilia. Downregulated expression of Anks3 in vivo in mice by Locked Nucleic Acid (LNA) modified antisense oligonucleotides was associated with increased transcription of vasopressin-induced genes, suggesting changes in renal water permeability, and altered transcription of genes encoding proteins involved in cilium structure, apoptosis and cell proliferation. These data provide experimental evidence of ANKS3-ANKS6 direct interaction through their SAM domain and co-localisation in mouse renal cilia, and shed light on molecular mechanisms indirectly mediated by ANKS6 in the mouse kidney, that may be affected by altered ANKS3-ANKS6 interaction. Our results contribute to improved knowledge of the structure and function of the network of proteins interacting with ANKS6, which may represent therapeutic targets in cystic diseases.

No MeSH data available.


Related in: MedlinePlus