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The cytokinesis gene KEULE encodes a Sec1 protein that binds the syntaxin KNOLLE.

Assaad FF, Huet Y, Mayer U, Jürgens G - J. Cell Biol. (2001)

Bottom Line: KEULE is characteristic of a Sec1 protein in that it appears to exist in two forms: soluble or peripherally associated with membranes.More importantly, KEULE binds the cytokinesis-specific syntaxin KNOLLE.Sec1 proteins are key regulators of vesicle trafficking, capable of integrating a large number of intra- and/or intercellular signals.

View Article: PubMed Central - PubMed

Affiliation: Genetics and Microbiology Institute, Ludwig Maximilians University, D-80638 Munich, Germany. fassaad@andrew2.stanford.edu

ABSTRACT
KEULE is required for cytokinesis in Arabidopsis thaliana. We have positionally cloned the KEULE gene and shown that it encodes a Sec1 protein. KEULE is expressed throughout the plant, yet appears enriched in dividing tissues. Cytokinesis-defective mutant sectors were observed in all somatic tissues upon transformation of wild-type plants with a KEULE-green fluorescent protein gene fusion, suggesting that KEULE is required not only during embryogenesis, but at all stages of the plant's life cycle. KEULE is characteristic of a Sec1 protein in that it appears to exist in two forms: soluble or peripherally associated with membranes. More importantly, KEULE binds the cytokinesis-specific syntaxin KNOLLE. Sec1 proteins are key regulators of vesicle trafficking, capable of integrating a large number of intra- and/or intercellular signals. As a cytokinesis-related Sec1 protein, KEULE appears to represent a novel link between cell cycle progression and the membrane fusion apparatus.

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Related in: MedlinePlus

KEULE antibody and expression of the KEULE protein visualized by Western analysis. (A) The peptide antibody specifically recognizes a 73-kD band in keule-like mutants (described in Materials and Methods) but no band of the size of KEULE in keule mutants. Bottom panel shows the dominant contaminating band revealed by this peptide antibody. Both lanes are loaded with 50 mutant seedlings (left, fast-neutron–induced keule allele; right, cytokinesis-defective line G235) homogenized in 10 μl of sample buffer. (B) KEULE is expressed throughout the plant (as seen after longer exposures of the blot; expression in root shown in Fig. 7 B and in seedling in Fig. 8 C) and appears enriched in dividing tissues of Arabidopsis, namely the root tips and inflorescence meristems, designated “flower” in the figure. The membrane was stained with Ponceau S to monitor loading.
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Figure 5: KEULE antibody and expression of the KEULE protein visualized by Western analysis. (A) The peptide antibody specifically recognizes a 73-kD band in keule-like mutants (described in Materials and Methods) but no band of the size of KEULE in keule mutants. Bottom panel shows the dominant contaminating band revealed by this peptide antibody. Both lanes are loaded with 50 mutant seedlings (left, fast-neutron–induced keule allele; right, cytokinesis-defective line G235) homogenized in 10 μl of sample buffer. (B) KEULE is expressed throughout the plant (as seen after longer exposures of the blot; expression in root shown in Fig. 7 B and in seedling in Fig. 8 C) and appears enriched in dividing tissues of Arabidopsis, namely the root tips and inflorescence meristems, designated “flower” in the figure. The membrane was stained with Ponceau S to monitor loading.

Mentions: We compared the sequences of six KEULE cDNAs with 7 kb genomic DNA encompassing the gene and the upstream region. Sequence analysis with web-based tools (Netstart, which predicts start sites for transcription) suggested that two cDNAs are full length and that the KEULE gene spans a genomic interval of 5 kb, including 20 introns (Fig. 2 D). Upstream of KEULE, the intergenic region is only 352 nucleotides long. The predicted transcription unit is 2.4 kb in length and encodes a protein of 667 amino acids. This predicted size was confirmed by Northern analysis and RT-PCR (Fig. 2 F and not shown), as well as Western analysis (see Fig. 5 A).


The cytokinesis gene KEULE encodes a Sec1 protein that binds the syntaxin KNOLLE.

Assaad FF, Huet Y, Mayer U, Jürgens G - J. Cell Biol. (2001)

KEULE antibody and expression of the KEULE protein visualized by Western analysis. (A) The peptide antibody specifically recognizes a 73-kD band in keule-like mutants (described in Materials and Methods) but no band of the size of KEULE in keule mutants. Bottom panel shows the dominant contaminating band revealed by this peptide antibody. Both lanes are loaded with 50 mutant seedlings (left, fast-neutron–induced keule allele; right, cytokinesis-defective line G235) homogenized in 10 μl of sample buffer. (B) KEULE is expressed throughout the plant (as seen after longer exposures of the blot; expression in root shown in Fig. 7 B and in seedling in Fig. 8 C) and appears enriched in dividing tissues of Arabidopsis, namely the root tips and inflorescence meristems, designated “flower” in the figure. The membrane was stained with Ponceau S to monitor loading.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195996&req=5

Figure 5: KEULE antibody and expression of the KEULE protein visualized by Western analysis. (A) The peptide antibody specifically recognizes a 73-kD band in keule-like mutants (described in Materials and Methods) but no band of the size of KEULE in keule mutants. Bottom panel shows the dominant contaminating band revealed by this peptide antibody. Both lanes are loaded with 50 mutant seedlings (left, fast-neutron–induced keule allele; right, cytokinesis-defective line G235) homogenized in 10 μl of sample buffer. (B) KEULE is expressed throughout the plant (as seen after longer exposures of the blot; expression in root shown in Fig. 7 B and in seedling in Fig. 8 C) and appears enriched in dividing tissues of Arabidopsis, namely the root tips and inflorescence meristems, designated “flower” in the figure. The membrane was stained with Ponceau S to monitor loading.
Mentions: We compared the sequences of six KEULE cDNAs with 7 kb genomic DNA encompassing the gene and the upstream region. Sequence analysis with web-based tools (Netstart, which predicts start sites for transcription) suggested that two cDNAs are full length and that the KEULE gene spans a genomic interval of 5 kb, including 20 introns (Fig. 2 D). Upstream of KEULE, the intergenic region is only 352 nucleotides long. The predicted transcription unit is 2.4 kb in length and encodes a protein of 667 amino acids. This predicted size was confirmed by Northern analysis and RT-PCR (Fig. 2 F and not shown), as well as Western analysis (see Fig. 5 A).

Bottom Line: KEULE is characteristic of a Sec1 protein in that it appears to exist in two forms: soluble or peripherally associated with membranes.More importantly, KEULE binds the cytokinesis-specific syntaxin KNOLLE.Sec1 proteins are key regulators of vesicle trafficking, capable of integrating a large number of intra- and/or intercellular signals.

View Article: PubMed Central - PubMed

Affiliation: Genetics and Microbiology Institute, Ludwig Maximilians University, D-80638 Munich, Germany. fassaad@andrew2.stanford.edu

ABSTRACT
KEULE is required for cytokinesis in Arabidopsis thaliana. We have positionally cloned the KEULE gene and shown that it encodes a Sec1 protein. KEULE is expressed throughout the plant, yet appears enriched in dividing tissues. Cytokinesis-defective mutant sectors were observed in all somatic tissues upon transformation of wild-type plants with a KEULE-green fluorescent protein gene fusion, suggesting that KEULE is required not only during embryogenesis, but at all stages of the plant's life cycle. KEULE is characteristic of a Sec1 protein in that it appears to exist in two forms: soluble or peripherally associated with membranes. More importantly, KEULE binds the cytokinesis-specific syntaxin KNOLLE. Sec1 proteins are key regulators of vesicle trafficking, capable of integrating a large number of intra- and/or intercellular signals. As a cytokinesis-related Sec1 protein, KEULE appears to represent a novel link between cell cycle progression and the membrane fusion apparatus.

Show MeSH
Related in: MedlinePlus