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The NC1/endostatin domain of Caenorhabditis elegans type XVIII collagen affects cell migration and axon guidance.

Ackley BD, Crew JR, Elamaa H, Pihlajaniemi T, Kuo CJ, Kramer JM - J. Cell Biol. (2001)

Bottom Line: The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system.In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain.These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.

ABSTRACT
Type XVIII collagen is a homotrimeric basement membrane molecule of unknown function, whose COOH-terminal NC1 domain contains endostatin (ES), a potent antiangiogenic agent. The Caenorhabditis elegans collagen XVIII homologue, cle-1, encodes three developmentally regulated protein isoforms expressed predominantly in neurons. The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system. Deletion of the cle-1 NC1 domain results in viable fertile animals that display multiple cell migration and axon guidance defects. Particular defects can be rescued by ectopic expression of the NC1 domain, which is shown to be capable of forming trimers. In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain. These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

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The C. elegans NC1 domain trimerizes in vitro. (A) Purified CLE-1 NC1 (left) and ES domains (right) were analyzed by denaturing gel electrophoresis and Western blotting, either before (−) or after (+) cross-linking with EGS. After cross-linking, the 40-kD NC1 monomer migrates at ∼120 kD, indicating that it exists as a trimer. The mobility of the ES domain does not shift after EGS treatment, indicating that it exists as a monomer. (B) Summary of mechanosensory neuron migration data. Defective mechanosensory neuron migrations are observed when the NC1 domain is removed by the cg120 deletion. These defects can be rescued by ectopic NC1 domain expression. In contrast, ectopic ES domain expression causes mechanosensory neuron migration defects in the wild-type background and fails to rescue the NC1 deletion defects. Expression of the association domain and hinge regions fused to GFP, rather than ES, does not rescue cg120 migration defects and does not cause defects in the wild-type background.
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Figure 8: The C. elegans NC1 domain trimerizes in vitro. (A) Purified CLE-1 NC1 (left) and ES domains (right) were analyzed by denaturing gel electrophoresis and Western blotting, either before (−) or after (+) cross-linking with EGS. After cross-linking, the 40-kD NC1 monomer migrates at ∼120 kD, indicating that it exists as a trimer. The mobility of the ES domain does not shift after EGS treatment, indicating that it exists as a monomer. (B) Summary of mechanosensory neuron migration data. Defective mechanosensory neuron migrations are observed when the NC1 domain is removed by the cg120 deletion. These defects can be rescued by ectopic NC1 domain expression. In contrast, ectopic ES domain expression causes mechanosensory neuron migration defects in the wild-type background and fails to rescue the NC1 deletion defects. Expression of the association domain and hinge regions fused to GFP, rather than ES, does not rescue cg120 migration defects and does not cause defects in the wild-type background.

Mentions: A major functional difference between type XVIII collagen NC1 and ES domains is the ability of NC1 to oligomerize (Sasaki et al. 1998; Kuo et al. 2001, this issue). Recombinant CLE-1 NC1 domains expressed in human embryonic kidney 293 cells were found to form trimers (Fig. 8 A), whereas CLE-1 ES domains remained monomeric. Thus, the C. elegans domains behave similarly to the mammalian domains in regards to oligomerization, suggesting that the functional differences between ectopically expressed CLE-1 NC1 and ES domains may reflect differences in their ability to oligomerize via the association domain.


The NC1/endostatin domain of Caenorhabditis elegans type XVIII collagen affects cell migration and axon guidance.

Ackley BD, Crew JR, Elamaa H, Pihlajaniemi T, Kuo CJ, Kramer JM - J. Cell Biol. (2001)

The C. elegans NC1 domain trimerizes in vitro. (A) Purified CLE-1 NC1 (left) and ES domains (right) were analyzed by denaturing gel electrophoresis and Western blotting, either before (−) or after (+) cross-linking with EGS. After cross-linking, the 40-kD NC1 monomer migrates at ∼120 kD, indicating that it exists as a trimer. The mobility of the ES domain does not shift after EGS treatment, indicating that it exists as a monomer. (B) Summary of mechanosensory neuron migration data. Defective mechanosensory neuron migrations are observed when the NC1 domain is removed by the cg120 deletion. These defects can be rescued by ectopic NC1 domain expression. In contrast, ectopic ES domain expression causes mechanosensory neuron migration defects in the wild-type background and fails to rescue the NC1 deletion defects. Expression of the association domain and hinge regions fused to GFP, rather than ES, does not rescue cg120 migration defects and does not cause defects in the wild-type background.
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Related In: Results  -  Collection

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Figure 8: The C. elegans NC1 domain trimerizes in vitro. (A) Purified CLE-1 NC1 (left) and ES domains (right) were analyzed by denaturing gel electrophoresis and Western blotting, either before (−) or after (+) cross-linking with EGS. After cross-linking, the 40-kD NC1 monomer migrates at ∼120 kD, indicating that it exists as a trimer. The mobility of the ES domain does not shift after EGS treatment, indicating that it exists as a monomer. (B) Summary of mechanosensory neuron migration data. Defective mechanosensory neuron migrations are observed when the NC1 domain is removed by the cg120 deletion. These defects can be rescued by ectopic NC1 domain expression. In contrast, ectopic ES domain expression causes mechanosensory neuron migration defects in the wild-type background and fails to rescue the NC1 deletion defects. Expression of the association domain and hinge regions fused to GFP, rather than ES, does not rescue cg120 migration defects and does not cause defects in the wild-type background.
Mentions: A major functional difference between type XVIII collagen NC1 and ES domains is the ability of NC1 to oligomerize (Sasaki et al. 1998; Kuo et al. 2001, this issue). Recombinant CLE-1 NC1 domains expressed in human embryonic kidney 293 cells were found to form trimers (Fig. 8 A), whereas CLE-1 ES domains remained monomeric. Thus, the C. elegans domains behave similarly to the mammalian domains in regards to oligomerization, suggesting that the functional differences between ectopically expressed CLE-1 NC1 and ES domains may reflect differences in their ability to oligomerize via the association domain.

Bottom Line: The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system.In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain.These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.

ABSTRACT
Type XVIII collagen is a homotrimeric basement membrane molecule of unknown function, whose COOH-terminal NC1 domain contains endostatin (ES), a potent antiangiogenic agent. The Caenorhabditis elegans collagen XVIII homologue, cle-1, encodes three developmentally regulated protein isoforms expressed predominantly in neurons. The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system. Deletion of the cle-1 NC1 domain results in viable fertile animals that display multiple cell migration and axon guidance defects. Particular defects can be rescued by ectopic expression of the NC1 domain, which is shown to be capable of forming trimers. In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain. These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

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Related in: MedlinePlus