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Drosophila p120catenin plays a supporting role in cell adhesion but is not an essential adherens junction component.

Myster SH, Cavallo R, Anderson CT, Fox DT, Peifer M - J. Cell Biol. (2003)

Bottom Line: We generated alleles of p120 and found that mutants are viable and fertile and have no substantial changes in junction structure or function.However, p120 mutations strongly enhance mutations in the genes encoding DE-cadherin or Armadillo, the beta-catenin homologue.Finally, we examined the localization of p120 during embryogenesis. p120 localizes to adherens junctions, but its localization there is less universal than that of core adherens junction proteins.

View Article: PubMed Central - PubMed

Affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

ABSTRACT
Cadherin-catenin complexes, localized to adherens junctions, are essential for cell-cell adhesion. One means of regulating adhesion is through the juxtamembrane domain of the cadherin cytoplasmic tail. This region is the binding site for p120, leading to the hypothesis that p120 is a key regulator of cell adhesion. p120 has also been suggested to regulate the GTPase Rho and to regulate transcription via its binding partner Kaiso. To test these hypothesized functions, we turned to Drosophila, which has only a single p120 family member. It localizes to adherens junctions and binds the juxtamembrane region of DE-cadherin (DE-cad). We generated alleles of p120 and found that mutants are viable and fertile and have no substantial changes in junction structure or function. However, p120 mutations strongly enhance mutations in the genes encoding DE-cadherin or Armadillo, the beta-catenin homologue. Finally, we examined the localization of p120 during embryogenesis. p120 localizes to adherens junctions, but its localization there is less universal than that of core adherens junction proteins. Together, these data suggest that p120 is an important positive modulator of adhesion but that it is not an essential core component of adherens junctions.

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Related in: MedlinePlus

p120308 is a  allele. (A) p120308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486. (B) The p120 mutants are mRNA s. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905, is a control. A DNA control confirmed we were examining mRNA.
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fig4: p120308 is a allele. (A) p120308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486. (B) The p120 mutants are mRNA s. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905, is a control. A DNA control confirmed we were examining mRNA.

Mentions: We thus turned to an unbiased approach to obtaining p120 mutants. The BDGP recently initiated a screen for P element insertions in new genomic regions (http://flypush.imgen.bcm.tmc.edu/pscreen/). One P element, KG01086, is ∼7 kb 3′ to p120, and 2 kb 5′ to the adjacent gene, CG17486 (Fig. 4 A). KG01086 is homozygous viable. We removed secondary P element insertions and other background mutations by replacing the other chromosomes and recombining off the left arm of the 2nd chromosome. We then mobilized KG01086 and screened for transposition to a nearby site or deletions beginning in the P element and extending into adjacent DNA (unpublished data). Three deletions affecting p120 were isolated. All remove the entire p120 coding sequence as assessed by PCR of genomic DNA across the region (Fig. 4 A; unpublished data), and two of the three do not affect adjacent genes (Fig. 4 A). We focused on one of these alleles, p120308. We verified that this allele is , since it does not make p120 mRNA as assessed by RT-PCR (Fig. 4 B) or stable p120 protein as assessed by immunoblotting with two different antisera (Fig. 2, A and B).


Drosophila p120catenin plays a supporting role in cell adhesion but is not an essential adherens junction component.

Myster SH, Cavallo R, Anderson CT, Fox DT, Peifer M - J. Cell Biol. (2003)

p120308 is a  allele. (A) p120308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486. (B) The p120 mutants are mRNA s. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905, is a control. A DNA control confirmed we were examining mRNA.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2172674&req=5

fig4: p120308 is a allele. (A) p120308 deletes the entire p120 coding region, but does not affect other genes. Schematic as in Fig. 1 A. Genomic DNA from single wild-type or homozygous mutant flies was PCR amplified using primer pairs from the indicated regions between LD05623 and CG17486. (B) The p120 mutants are mRNA s. cDNA generated from oligo-dT-primed total RNA from p120 mutants and wild-type was amplified with primers spanning the p120 third intron. An unrelated gene, CG2905, is a control. A DNA control confirmed we were examining mRNA.
Mentions: We thus turned to an unbiased approach to obtaining p120 mutants. The BDGP recently initiated a screen for P element insertions in new genomic regions (http://flypush.imgen.bcm.tmc.edu/pscreen/). One P element, KG01086, is ∼7 kb 3′ to p120, and 2 kb 5′ to the adjacent gene, CG17486 (Fig. 4 A). KG01086 is homozygous viable. We removed secondary P element insertions and other background mutations by replacing the other chromosomes and recombining off the left arm of the 2nd chromosome. We then mobilized KG01086 and screened for transposition to a nearby site or deletions beginning in the P element and extending into adjacent DNA (unpublished data). Three deletions affecting p120 were isolated. All remove the entire p120 coding sequence as assessed by PCR of genomic DNA across the region (Fig. 4 A; unpublished data), and two of the three do not affect adjacent genes (Fig. 4 A). We focused on one of these alleles, p120308. We verified that this allele is , since it does not make p120 mRNA as assessed by RT-PCR (Fig. 4 B) or stable p120 protein as assessed by immunoblotting with two different antisera (Fig. 2, A and B).

Bottom Line: We generated alleles of p120 and found that mutants are viable and fertile and have no substantial changes in junction structure or function.However, p120 mutations strongly enhance mutations in the genes encoding DE-cadherin or Armadillo, the beta-catenin homologue.Finally, we examined the localization of p120 during embryogenesis. p120 localizes to adherens junctions, but its localization there is less universal than that of core adherens junction proteins.

View Article: PubMed Central - PubMed

Affiliation: Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

ABSTRACT
Cadherin-catenin complexes, localized to adherens junctions, are essential for cell-cell adhesion. One means of regulating adhesion is through the juxtamembrane domain of the cadherin cytoplasmic tail. This region is the binding site for p120, leading to the hypothesis that p120 is a key regulator of cell adhesion. p120 has also been suggested to regulate the GTPase Rho and to regulate transcription via its binding partner Kaiso. To test these hypothesized functions, we turned to Drosophila, which has only a single p120 family member. It localizes to adherens junctions and binds the juxtamembrane region of DE-cadherin (DE-cad). We generated alleles of p120 and found that mutants are viable and fertile and have no substantial changes in junction structure or function. However, p120 mutations strongly enhance mutations in the genes encoding DE-cadherin or Armadillo, the beta-catenin homologue. Finally, we examined the localization of p120 during embryogenesis. p120 localizes to adherens junctions, but its localization there is less universal than that of core adherens junction proteins. Together, these data suggest that p120 is an important positive modulator of adhesion but that it is not an essential core component of adherens junctions.

Show MeSH
Related in: MedlinePlus