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Drosophila roadblock and Chlamydomonas LC7: a conserved family of dynein-associated proteins involved in axonal transport, flagellar motility, and mitosis.

Bowman AB, Patel-King RS, Benashski SE, McCaffery JM, Goldstein LS, King SM - J. Cell Biol. (1999)

Bottom Line: The gene identified by robl encodes a 97-amino acid polypeptide that is 57% identical (70% similar) to the 105-amino acid Chlamydomonas outer arm dynein-associated protein LC7, also reported here.Furthermore, we demonstrate that members of this family of proteins are associated with both flagellar outer arm dynein and Drosophila and rat brain cytoplasmic dynein.We propose that roadblock/LC7 family members may modulate specific dynein functions.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Division of Cellular and Molecular Medicine, Department of Pharmacology, University of California San Diego, La Jolla, California 92093-0683, USA.

ABSTRACT
Eukaryotic organisms utilize microtubule-dependent motors of the kinesin and dynein superfamilies to generate intracellular movement. To identify new genes involved in the regulation of axonal transport in Drosophila melanogaster, we undertook a screen based upon the sluggish larval phenotype of known motor mutants. One of the mutants identified in this screen, roadblock (robl), exhibits diverse defects in intracellular transport including axonal transport and mitosis. These defects include intra-axonal accumulations of cargoes, severe axonal degeneration, and aberrant chromosome segregation. The gene identified by robl encodes a 97-amino acid polypeptide that is 57% identical (70% similar) to the 105-amino acid Chlamydomonas outer arm dynein-associated protein LC7, also reported here. Both robl and LC7 have homology to several other genes from fruit fly, nematode, and mammals, but not Saccharomyces cerevisiae. Furthermore, we demonstrate that members of this family of proteins are associated with both flagellar outer arm dynein and Drosophila and rat brain cytoplasmic dynein. We propose that roadblock/LC7 family members may modulate specific dynein functions.

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A large family of robl/LC7-like proteins. BLAST analysis has identified several mammalian ESTs, Drosophila genes, and a gene from C. elegans that are highly homologous to robl/LC7. (A) A dendrogram of the robl/LC7-like family members identified is shown. This dendrogram was generated using the GCG pileup command. We identified at least five Drosophila roadblock-like genes by searching the BDGP-derived ESTs and genomic sequences. Previously unidentified genes have been designated by their cytological location determined by BDGP. Also, two classes of robl/LC7-like genes have been identified as mammalian ESTs (identified by a representative EST accession number). (B) An alignment of the protein family is shown. The alignment was generated by the same GCG pileup command as an MSF file. Boxshade was used to illustrate aligned amino acid identity (dark shaded residues) and similarity (light shaded residues).
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Figure 4: A large family of robl/LC7-like proteins. BLAST analysis has identified several mammalian ESTs, Drosophila genes, and a gene from C. elegans that are highly homologous to robl/LC7. (A) A dendrogram of the robl/LC7-like family members identified is shown. This dendrogram was generated using the GCG pileup command. We identified at least five Drosophila roadblock-like genes by searching the BDGP-derived ESTs and genomic sequences. Previously unidentified genes have been designated by their cytological location determined by BDGP. Also, two classes of robl/LC7-like genes have been identified as mammalian ESTs (identified by a representative EST accession number). (B) An alignment of the protein family is shown. The alignment was generated by the same GCG pileup command as an MSF file. Boxshade was used to illustrate aligned amino acid identity (dark shaded residues) and similarity (light shaded residues).

Mentions: All sequence assembly and protein comparison were performed using the GCG suite of software (Genetics Computer Group) and Sequencher 3.1 software (Gene Codes Corporation). Roadblock/LC7 family members were identified with the Drosophila robl sequence using BLASTP against the NCBI dbNR, tblastn against the NCBI dbEST, and tblastn against the BDGP DNA sequence database. All gene abbreviations here refer to those detailed in Fig. 4. Drosophila robl22E (DS01020; accession number AC004276), robl37BC (DS00790; accession number AC005127), robl62A (DS02734; accession number AC004343), and robl60C (DS02336; accession number AC005718) were identified in BDGP genomic sequence and are apparently intronless genes, just like the related late RNA from the Bithoraxoid complex (accession number M27999). EST clones have been identified for late RNA-encoded bithoraxoid protein (bxd) (GH08635; accession number AI113381) and robl62A (GH15530; accession number AI292590) by BDGP from a Drosophila head cDNA library. The proteins T24H10.6 (accession number 995857) and bxd (accession number 290293) were identified from dbNR using blastp. Mouse, rat, and human ESTs identified, were compared by nucleotide sequence using tblastn against the species-specific NCBI GenBank dbEST to identify ESTs from identical genes; two different genes were identified in all three species (accession numbers from representative ESTs are given in Fig. 4). The predicted translation of all mammalian ESTs was determined using DNA Strider (CEA); the small size of the genes meant that almost all ESTs translated into full-length protein. Protein comparison was done using the GCG pileup command to generate the dendrogram; the output MSF file was run through the BoxShade Server (http://www. isrec.isb-sib.ch/software/BOX_form.html) and the output EPS file was imported into Adobe Illustrator 6.0.


Drosophila roadblock and Chlamydomonas LC7: a conserved family of dynein-associated proteins involved in axonal transport, flagellar motility, and mitosis.

Bowman AB, Patel-King RS, Benashski SE, McCaffery JM, Goldstein LS, King SM - J. Cell Biol. (1999)

A large family of robl/LC7-like proteins. BLAST analysis has identified several mammalian ESTs, Drosophila genes, and a gene from C. elegans that are highly homologous to robl/LC7. (A) A dendrogram of the robl/LC7-like family members identified is shown. This dendrogram was generated using the GCG pileup command. We identified at least five Drosophila roadblock-like genes by searching the BDGP-derived ESTs and genomic sequences. Previously unidentified genes have been designated by their cytological location determined by BDGP. Also, two classes of robl/LC7-like genes have been identified as mammalian ESTs (identified by a representative EST accession number). (B) An alignment of the protein family is shown. The alignment was generated by the same GCG pileup command as an MSF file. Boxshade was used to illustrate aligned amino acid identity (dark shaded residues) and similarity (light shaded residues).
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Figure 4: A large family of robl/LC7-like proteins. BLAST analysis has identified several mammalian ESTs, Drosophila genes, and a gene from C. elegans that are highly homologous to robl/LC7. (A) A dendrogram of the robl/LC7-like family members identified is shown. This dendrogram was generated using the GCG pileup command. We identified at least five Drosophila roadblock-like genes by searching the BDGP-derived ESTs and genomic sequences. Previously unidentified genes have been designated by their cytological location determined by BDGP. Also, two classes of robl/LC7-like genes have been identified as mammalian ESTs (identified by a representative EST accession number). (B) An alignment of the protein family is shown. The alignment was generated by the same GCG pileup command as an MSF file. Boxshade was used to illustrate aligned amino acid identity (dark shaded residues) and similarity (light shaded residues).
Mentions: All sequence assembly and protein comparison were performed using the GCG suite of software (Genetics Computer Group) and Sequencher 3.1 software (Gene Codes Corporation). Roadblock/LC7 family members were identified with the Drosophila robl sequence using BLASTP against the NCBI dbNR, tblastn against the NCBI dbEST, and tblastn against the BDGP DNA sequence database. All gene abbreviations here refer to those detailed in Fig. 4. Drosophila robl22E (DS01020; accession number AC004276), robl37BC (DS00790; accession number AC005127), robl62A (DS02734; accession number AC004343), and robl60C (DS02336; accession number AC005718) were identified in BDGP genomic sequence and are apparently intronless genes, just like the related late RNA from the Bithoraxoid complex (accession number M27999). EST clones have been identified for late RNA-encoded bithoraxoid protein (bxd) (GH08635; accession number AI113381) and robl62A (GH15530; accession number AI292590) by BDGP from a Drosophila head cDNA library. The proteins T24H10.6 (accession number 995857) and bxd (accession number 290293) were identified from dbNR using blastp. Mouse, rat, and human ESTs identified, were compared by nucleotide sequence using tblastn against the species-specific NCBI GenBank dbEST to identify ESTs from identical genes; two different genes were identified in all three species (accession numbers from representative ESTs are given in Fig. 4). The predicted translation of all mammalian ESTs was determined using DNA Strider (CEA); the small size of the genes meant that almost all ESTs translated into full-length protein. Protein comparison was done using the GCG pileup command to generate the dendrogram; the output MSF file was run through the BoxShade Server (http://www. isrec.isb-sib.ch/software/BOX_form.html) and the output EPS file was imported into Adobe Illustrator 6.0.

Bottom Line: The gene identified by robl encodes a 97-amino acid polypeptide that is 57% identical (70% similar) to the 105-amino acid Chlamydomonas outer arm dynein-associated protein LC7, also reported here.Furthermore, we demonstrate that members of this family of proteins are associated with both flagellar outer arm dynein and Drosophila and rat brain cytoplasmic dynein.We propose that roadblock/LC7 family members may modulate specific dynein functions.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Division of Cellular and Molecular Medicine, Department of Pharmacology, University of California San Diego, La Jolla, California 92093-0683, USA.

ABSTRACT
Eukaryotic organisms utilize microtubule-dependent motors of the kinesin and dynein superfamilies to generate intracellular movement. To identify new genes involved in the regulation of axonal transport in Drosophila melanogaster, we undertook a screen based upon the sluggish larval phenotype of known motor mutants. One of the mutants identified in this screen, roadblock (robl), exhibits diverse defects in intracellular transport including axonal transport and mitosis. These defects include intra-axonal accumulations of cargoes, severe axonal degeneration, and aberrant chromosome segregation. The gene identified by robl encodes a 97-amino acid polypeptide that is 57% identical (70% similar) to the 105-amino acid Chlamydomonas outer arm dynein-associated protein LC7, also reported here. Both robl and LC7 have homology to several other genes from fruit fly, nematode, and mammals, but not Saccharomyces cerevisiae. Furthermore, we demonstrate that members of this family of proteins are associated with both flagellar outer arm dynein and Drosophila and rat brain cytoplasmic dynein. We propose that roadblock/LC7 family members may modulate specific dynein functions.

Show MeSH
Related in: MedlinePlus