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Hedgehog signal transduction proteins: contacts of the Fused kinase and Ci transcription factor with the kinesin-related protein Costal2.

Monnier V, Ho KS, Sanial M, Scott MP, Plessis A - BMC Dev. Biol. (2002)

Bottom Line: In flies, key Hedgehog signal transduction components have been identified including the kinesin-related protein Costal2, the serinethreonine kinase Fused, and the PEST-containing protein Suppressor of Fused.In fly embryos, Costal2, Fused, Suppressor of Fused and Cubitus interruptus are associated in at least one cytoplasmic complex, which interacts with the microtubules in a Hedgehog-dependent manner.Our results provide new insights into the possible mechanism of the cytosolic steps of Hedgehog transduction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de génétique et évolution, Institut Jacques Monod, 2 Place Jussieu, 75005, Paris, France. monnier@ijm.jussieu.fr

ABSTRACT

Background: Hedgehog signaling proteins play important roles in development by controlling growth and patterning in various animals including Drosophila and mammals. Hedgehog signaling triggers changes in responsive cells through a novel transduction mechanism that ultimately controls the transcription of specific target genes via the activity of zinc finger transcription factors of the Cubitus interruptus/GLI family. In flies, key Hedgehog signal transduction components have been identified including the kinesin-related protein Costal2, the serinethreonine kinase Fused, and the PEST-containing protein Suppressor of Fused. These proteins control Cubitus interruptus cleavage, nucleo-cytoplasmic localization and activation. In fly embryos, Costal2, Fused, Suppressor of Fused and Cubitus interruptus are associated in at least one cytoplasmic complex, which interacts with the microtubules in a Hedgehog-dependent manner.

Results: Here we identified and mapped direct interactions between Cos2, Fu, and Ci using an in vitro affinity assay and the yeast two-hybrid system.

Conclusions: Our results provide new insights into the possible mechanism of the cytosolic steps of Hedgehog transduction.

Show MeSH
Summary of the interaction domains The interactions between Cos2, Ci, Fu, and Fureg described above and in former work [31] are summarized. The interaction between the region 941–1065 of Ci and Cos2 was previously shown by Wang et al. 2000 [23]. The domains of interactions are indicated in lines above or under each protein structure. No interaction could be detected between Cos2 and Sufu (data not shown). See also legends for Figures 1 and 2. M: Cos2 motor domain, HR: Cos2 heptad repeats, DBD:Ci DNA binding domain; NLS: Ci nuclear localisation signal, AD: Ci activation domain.
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Figure 3: Summary of the interaction domains The interactions between Cos2, Ci, Fu, and Fureg described above and in former work [31] are summarized. The interaction between the region 941–1065 of Ci and Cos2 was previously shown by Wang et al. 2000 [23]. The domains of interactions are indicated in lines above or under each protein structure. No interaction could be detected between Cos2 and Sufu (data not shown). See also legends for Figures 1 and 2. M: Cos2 motor domain, HR: Cos2 heptad repeats, DBD:Ci DNA binding domain; NLS: Ci nuclear localisation signal, AD: Ci activation domain.

Mentions: In the yeast two-hybrid assay, Fureg (Fu306-805) interacts specifically with the C-terminal part of Cos2 (Cos538-1201) (Figure 1B). Part of this region, Cos538-751, is sufficient to interact with Fureg. Cos538-751 encodes a region of Cos2 C-terminal to the putative motor domain (See also Figure 3) and which overlaps 14 of the 36 heptad repeats of a predicted coiled-coil motif of Cos2. In other kinesins the coiled coil repeats mediate homodimerization and belong to a flexible stalk domain involved in the movements of the motor domain along the microtubules [34-45].


Hedgehog signal transduction proteins: contacts of the Fused kinase and Ci transcription factor with the kinesin-related protein Costal2.

Monnier V, Ho KS, Sanial M, Scott MP, Plessis A - BMC Dev. Biol. (2002)

Summary of the interaction domains The interactions between Cos2, Ci, Fu, and Fureg described above and in former work [31] are summarized. The interaction between the region 941–1065 of Ci and Cos2 was previously shown by Wang et al. 2000 [23]. The domains of interactions are indicated in lines above or under each protein structure. No interaction could be detected between Cos2 and Sufu (data not shown). See also legends for Figures 1 and 2. M: Cos2 motor domain, HR: Cos2 heptad repeats, DBD:Ci DNA binding domain; NLS: Ci nuclear localisation signal, AD: Ci activation domain.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC101406&req=5

Figure 3: Summary of the interaction domains The interactions between Cos2, Ci, Fu, and Fureg described above and in former work [31] are summarized. The interaction between the region 941–1065 of Ci and Cos2 was previously shown by Wang et al. 2000 [23]. The domains of interactions are indicated in lines above or under each protein structure. No interaction could be detected between Cos2 and Sufu (data not shown). See also legends for Figures 1 and 2. M: Cos2 motor domain, HR: Cos2 heptad repeats, DBD:Ci DNA binding domain; NLS: Ci nuclear localisation signal, AD: Ci activation domain.
Mentions: In the yeast two-hybrid assay, Fureg (Fu306-805) interacts specifically with the C-terminal part of Cos2 (Cos538-1201) (Figure 1B). Part of this region, Cos538-751, is sufficient to interact with Fureg. Cos538-751 encodes a region of Cos2 C-terminal to the putative motor domain (See also Figure 3) and which overlaps 14 of the 36 heptad repeats of a predicted coiled-coil motif of Cos2. In other kinesins the coiled coil repeats mediate homodimerization and belong to a flexible stalk domain involved in the movements of the motor domain along the microtubules [34-45].

Bottom Line: In flies, key Hedgehog signal transduction components have been identified including the kinesin-related protein Costal2, the serinethreonine kinase Fused, and the PEST-containing protein Suppressor of Fused.In fly embryos, Costal2, Fused, Suppressor of Fused and Cubitus interruptus are associated in at least one cytoplasmic complex, which interacts with the microtubules in a Hedgehog-dependent manner.Our results provide new insights into the possible mechanism of the cytosolic steps of Hedgehog transduction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de génétique et évolution, Institut Jacques Monod, 2 Place Jussieu, 75005, Paris, France. monnier@ijm.jussieu.fr

ABSTRACT

Background: Hedgehog signaling proteins play important roles in development by controlling growth and patterning in various animals including Drosophila and mammals. Hedgehog signaling triggers changes in responsive cells through a novel transduction mechanism that ultimately controls the transcription of specific target genes via the activity of zinc finger transcription factors of the Cubitus interruptus/GLI family. In flies, key Hedgehog signal transduction components have been identified including the kinesin-related protein Costal2, the serinethreonine kinase Fused, and the PEST-containing protein Suppressor of Fused. These proteins control Cubitus interruptus cleavage, nucleo-cytoplasmic localization and activation. In fly embryos, Costal2, Fused, Suppressor of Fused and Cubitus interruptus are associated in at least one cytoplasmic complex, which interacts with the microtubules in a Hedgehog-dependent manner.

Results: Here we identified and mapped direct interactions between Cos2, Fu, and Ci using an in vitro affinity assay and the yeast two-hybrid system.

Conclusions: Our results provide new insights into the possible mechanism of the cytosolic steps of Hedgehog transduction.

Show MeSH