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The Drosophila CD2AP/CIN85 orthologue Cindr regulates junctions and cytoskeleton dynamics during tissue patterning.

Johnson RI, Seppa MJ, Cagan RL - J. Cell Biol. (2008)

Bottom Line: Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death.Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta.Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental and Regenerative Biology, Mount Sinai Medical School, New York, NY 10029, USA.

ABSTRACT
Developing tissues require cells to undergo intricate processes to shift into appropriate niches. This requires a functional connection between adhesion-mediating events at the cell surface and a cytoskeletal reorganization to permit directed movement. A small number of proteins are proposed to link these processes. Here, we identify one candidate, Cindr, the sole Drosophila melanogaster member of the CD2AP/CIN85 family (this family has been previously implicated in a variety of processes). Using D. melanogaster retina, we demonstrate that Cindr links cell surface junctions (E-cadherin) and adhesion (Roughest) with multiple components of the actin cytoskeleton. Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death. Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta. Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

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cindr interacts with cpb and cpa. (A) GMR>cindr-IR2 retina probed for DE-Cad (green, A′) and Rst (red, A″). (B) cpbF-19/GMR>cindr-IR2. Mispatterning was strongly enhanced. (C) cpbbnd1/GMR>cindr-IR2. (D) In wild-type retinae, Cpb (green) and Cindr (blue) colocalized. DE-Cad is shown in red. (E) Western analysis of final eluants of coimmunopreciptations from embryos expressing cindr-PCTAP or TAP only probed with α-Cindr (top) or α-Cpb (bottom). (F–H) cindr-IR2 was strongly enhanced in trans to cpa heterozygotes or when cpa was overexpressed. (F) cpascrd1/GMR>cindr-IR2. (G) cpa69E/GMR>cindr-IR2. (H) GMR>cindr-IR2, cpa. All tissue was dissected at 41 h APF; DE-Cad is only shown in C, F, G, and H. Bars, 10 μm.
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fig6: cindr interacts with cpb and cpa. (A) GMR>cindr-IR2 retina probed for DE-Cad (green, A′) and Rst (red, A″). (B) cpbF-19/GMR>cindr-IR2. Mispatterning was strongly enhanced. (C) cpbbnd1/GMR>cindr-IR2. (D) In wild-type retinae, Cpb (green) and Cindr (blue) colocalized. DE-Cad is shown in red. (E) Western analysis of final eluants of coimmunopreciptations from embryos expressing cindr-PCTAP or TAP only probed with α-Cindr (top) or α-Cpb (bottom). (F–H) cindr-IR2 was strongly enhanced in trans to cpa heterozygotes or when cpa was overexpressed. (F) cpascrd1/GMR>cindr-IR2. (G) cpa69E/GMR>cindr-IR2. (H) GMR>cindr-IR2, cpa. All tissue was dissected at 41 h APF; DE-Cad is only shown in C, F, G, and H. Bars, 10 μm.

Mentions: CPα/CPβ heterodimers reversibly bind barbed, fast-growing ends of actin filaments to prevent further elongation or depolymerization (Isenberg et al., 1980; Hopmann et al., 1996). Demonstrating a functional link between cindr and the D. melanogaster orthologues cpa/cpb, we found that multiple alleles of either cpa or cpb strongly and dominantly enhanced the patterning defects of GMR>cindr-IR2 (Figs. 2 and 5, B, C, F, and G). Similarly, we observed an enhancement of the cindr-IR phenotype when ectopic cpa or cpb was concurrently expressed (Fig. 6 H and not depicted); neither protein gave rise to a detectable phenotype when misexpressed alone (not depicted). The ability of both increased and decreased cpa/cpb activity to enhance cindr-dependent phenotypes indicates that the balance of Cindr and Cpa/Cpb is important. Cpb protein was expressed in all cell types within the pupal eye (Fig. 6 D) and the distribution and levels of Cpb were not detectably altered in a cindr-IR context (not depicted).


The Drosophila CD2AP/CIN85 orthologue Cindr regulates junctions and cytoskeleton dynamics during tissue patterning.

Johnson RI, Seppa MJ, Cagan RL - J. Cell Biol. (2008)

cindr interacts with cpb and cpa. (A) GMR>cindr-IR2 retina probed for DE-Cad (green, A′) and Rst (red, A″). (B) cpbF-19/GMR>cindr-IR2. Mispatterning was strongly enhanced. (C) cpbbnd1/GMR>cindr-IR2. (D) In wild-type retinae, Cpb (green) and Cindr (blue) colocalized. DE-Cad is shown in red. (E) Western analysis of final eluants of coimmunopreciptations from embryos expressing cindr-PCTAP or TAP only probed with α-Cindr (top) or α-Cpb (bottom). (F–H) cindr-IR2 was strongly enhanced in trans to cpa heterozygotes or when cpa was overexpressed. (F) cpascrd1/GMR>cindr-IR2. (G) cpa69E/GMR>cindr-IR2. (H) GMR>cindr-IR2, cpa. All tissue was dissected at 41 h APF; DE-Cad is only shown in C, F, G, and H. Bars, 10 μm.
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fig6: cindr interacts with cpb and cpa. (A) GMR>cindr-IR2 retina probed for DE-Cad (green, A′) and Rst (red, A″). (B) cpbF-19/GMR>cindr-IR2. Mispatterning was strongly enhanced. (C) cpbbnd1/GMR>cindr-IR2. (D) In wild-type retinae, Cpb (green) and Cindr (blue) colocalized. DE-Cad is shown in red. (E) Western analysis of final eluants of coimmunopreciptations from embryos expressing cindr-PCTAP or TAP only probed with α-Cindr (top) or α-Cpb (bottom). (F–H) cindr-IR2 was strongly enhanced in trans to cpa heterozygotes or when cpa was overexpressed. (F) cpascrd1/GMR>cindr-IR2. (G) cpa69E/GMR>cindr-IR2. (H) GMR>cindr-IR2, cpa. All tissue was dissected at 41 h APF; DE-Cad is only shown in C, F, G, and H. Bars, 10 μm.
Mentions: CPα/CPβ heterodimers reversibly bind barbed, fast-growing ends of actin filaments to prevent further elongation or depolymerization (Isenberg et al., 1980; Hopmann et al., 1996). Demonstrating a functional link between cindr and the D. melanogaster orthologues cpa/cpb, we found that multiple alleles of either cpa or cpb strongly and dominantly enhanced the patterning defects of GMR>cindr-IR2 (Figs. 2 and 5, B, C, F, and G). Similarly, we observed an enhancement of the cindr-IR phenotype when ectopic cpa or cpb was concurrently expressed (Fig. 6 H and not depicted); neither protein gave rise to a detectable phenotype when misexpressed alone (not depicted). The ability of both increased and decreased cpa/cpb activity to enhance cindr-dependent phenotypes indicates that the balance of Cindr and Cpa/Cpb is important. Cpb protein was expressed in all cell types within the pupal eye (Fig. 6 D) and the distribution and levels of Cpb were not detectably altered in a cindr-IR context (not depicted).

Bottom Line: Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death.Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta.Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental and Regenerative Biology, Mount Sinai Medical School, New York, NY 10029, USA.

ABSTRACT
Developing tissues require cells to undergo intricate processes to shift into appropriate niches. This requires a functional connection between adhesion-mediating events at the cell surface and a cytoskeletal reorganization to permit directed movement. A small number of proteins are proposed to link these processes. Here, we identify one candidate, Cindr, the sole Drosophila melanogaster member of the CD2AP/CIN85 family (this family has been previously implicated in a variety of processes). Using D. melanogaster retina, we demonstrate that Cindr links cell surface junctions (E-cadherin) and adhesion (Roughest) with multiple components of the actin cytoskeleton. Reducing cindr activity leads to defects in local cell movement and, consequently, tissue patterning and cell death. Cindr activity is required for normal localization of Drosophila E-cadherin and Roughest, and we show additional physical and functional links to multiple components of the actin cytoskeleton, including the actin-capping proteins capping protein alpha and capping protein beta. Together, these data demonstrate that Cindr is involved in dynamic cell rearrangement in an emerging epithelium.

Show MeSH
Related in: MedlinePlus