Hueston JL , Herren GP , Cueva JG , Buechner M , Lundquist EA , Goodman MB , Suprenant KA .

R01 NS047715-05A2 NINDS NIH HHS , R01 NS040945 NINDS NIH HHS , DK 55526 NIDDK NIH HHS , NS 40945 NINDS NIH HHS , T32 GM008545 NIGMS NIH HHS , R56 NS040945 NINDS NIH HHS , NS 047715 NINDS NIH HHS , R01 DK055526 NIDDK NIH HHS , R01 NS047715 NINDS NIH HHS , GM 08545 NIGMS NIH HHS

	Figure 1. ELP-1 domain structure. All members of the EMAP-like protein family are constructed with a Hydrophobic ELP (HELP) motif (PF03451) preceding a WD domain (PF00400). A COILED domain (PF05710) is predicted in C. elegans ELP-1 as well as in three of the five human EMLs. An additional doublecortin (DCX) domain (PF 03607) is predicted in fruit fly ELP-1. Accession numbers for the domains/motifs (parentheses) are provided from the Protein Family (PFAM) database [57]. The accession numbers for the human EMLs are as follows: EML1 [GenBank: NM_001008707], EML2 [GenBank: NM_012155], EML3 [GenBank: NM_153265], EML4 [GenBank: NM_019063], and EML5 [GenBank: NM_183387].
	Figure 2. Genetic and molecular characterization of elp-1. (A) The physical map of elp-1 is drawn in the reverse orientation. Two alternatively spliced transcripts, elp-1a and elp-1b, are shown with exons as solid boxes and introns as lines. We confirmed the identity of the PCR products by a diagnostic restriction digest with EcoR1 and BglII (Panel A, right). (B) Genetic structures of the elp-1::gfp expression constructs pKA99-1, pKA99-2 and pKA99-3. Exons are indicated by black boxes. The proteins encoded by these constructs are indicated as Pelp-1::GFP, ELP- 1Δ::NLS::GFP, and ELP-1::GFP, respectively in the Text. (C) Amino acid sequence of ELP-1a showing a potential alternative start site for elp-1(ok347) (black arrow), exon 5 aa sequence (magenta), HELP motif (green) and the beginning of the WD domain (red arrow). (D) PCR reaction with a forward primer in the deletion site and a reverse primer in exon 11. There is no detectable wild-type copy of the elp-1 gene in the deletion strain as evidenced by the absence of the 1105 bp band. (E) The elp-1(ok347) strain retains two mRNAs of approximately 2200 and 2100 bp.
	Figure 3. ELP-1::GFP is expressed in hypodermal cells at the 1 1/2-fold stage of development. Two focal planes within the same embryo are examined by differential interference contrast (DIC) (A, C) and fluorescent light microscopy (B, D). The bar represents 10 μm.
	Figure 5. ELP-1 is expressed in diverse muscle cellular junctions. (A) A triplet of dots surrounds the fusiform-shaped body wall muscle cell. ELP-1::GFP is located throughout the muscle arms (arrowheads) that make junctions with the ventral nerve cord. The nucleus is shown in this plane of focus. (B) ELP-1::GFP is located at the adhesion sites of the male-specific sex muscles (boxed insert). The auto-fluorescent spicules of the male tail are visible. A body wall muscle cell is observed below the sex-muscle adhesion sites. The cell is outlined with triplet dots. Dotted arrows show a linear expression pattern in this body wall muscle cell. Bar represents 10 μm.
	Figure 6. ELP-1 localizes to body wall muscle in hermaphrodites. (A) Near the muscle cell membrane, ELP-1::GFP is prominent in lines (small arrows) oriented with the longitudinal axis of the cell. ELP-1::GFP is also associated with an array of fluorescent filaments (jagged arrow). (B) Affinity-purified antibodies against ELP-1 also stain a linear array near the cell surface. Bar represents 10 μm. Within these "lines" there often is a repeating unit of fluorescence that is apparent in both transgenic animals (C) and in antibody-stained animals (D). Bar represents 2.5 μm. (E) Dense bodies (arrowheads) are shown at the muscle cell surface by phase contrast microscopy. ELP-1::GFP expression was examined via fluorescence light microscopy (F) and the two images taken at the same focal plane were overlaid in Panel G. Bar represents 1 μm. Muscle cells that were double-stained with antibodies against ELP-1 (H, J) and PAT-3/β-integrin (MH25) (I, and J) show overlapping staining patterns. The integrin puncta are superimposed upon the more linear staining pattern of ELP-1. When the membrane is removed by the freeze-cracking procedure (outlined in white in panel J), integrin is lost and the ELP-1 antigen remains. Bar represents 10 μm.
	Figure 7. Analysis of subcellular filaments in a single body wall muscle cell of a hermaphrodite. These three panels (A-C) were part of a Z-series of images that were taken every 0.2 μm from the muscle cell membrane towards the interior of the muscle cell. The first image in Panel A was taken approximately 0.8 μm from the cell surface (4 sections down from the surface). At this focal plane, ELP-1::GFP is associated with oblique linear striations overlap with the anti-β integrin (MH25) staining pattern shown in Figure 6. In successive focal planes (B and C) the filaments are no longer in linear arrays and have branched off into a criss-crossing array. Bar represents 10 μm.
	Figure 9. ELP-1 antigens co-pellet with paclitaxel-stabilized C. elegans microtubules. Paclitaxel-stabilized microtubules (Mt) were prepared from a mixed-stage worm extract (Ex) as described in the Materials and Methods. The panel on the left shows the protein complexity in a C. elegans extract (Ex, 25,000 g supernatant) and in a Paclitaxel-stabilized microtubule preparation (Mt, 25,000 g pellet). An ~100 kDa polypeptide and α and β tubulin are the most prominent proteins visualized in the microtubule preparation. Proteins were separated on an 8% acrylamide mini-gel (left panel) and probed with affinity-purified anti-ELP-1 antibodies on a nitrocellulose blot (right panel). Asterisk is next to the ~100 kDa band that cross-reacts with the anti-ELP-1 antibodies. Molecular masses × 10-3 (Mr) are shown to the left. Approximately 15 μg of protein was loaded in each lane of a 7% polyacrylamide SDS-page mini-gel.
	Figure 10. ELP-1 is needed for normal touch sensation. Bars are mean ± s.d. for at least three independent assays of 30–50 animals each. RNAi knockdown of elp-1 in the lin-15b;eri-1 background induces a stronger touch-insensitive phenotype (66 ± 11%, mean ± s.d.) than does partial deletion of elp-1(ok347) (24 ± 5%) and is similar to the defect found in ok347/ozDf1 heterozygotes (82 ± 6%). Percentage of touch-insensitive animals for the remaining genotypes and RNAi treatments are as follows: wild-type (12 ± 0%) and lin-15b;eri-1 without RNAi (19 ± 6%). Tests were conducted blind to genotype or RNAi treatment. *P < 0.001 Fisher's Exact test.
	Figure 11. Three-dimensional sketch illustrating the relationship of ELP-1-associated microtubules and the dense bodies (green). A transverse section through a single body wall muscle cell is shown. The thin filaments, which are attached to the dense bodies, and the thick filaments attached to the M-lines, have been eliminated from the sketch for clarity. ELP-1 associated microtubules (magenta) are associated with the apical portion of the dense body. Neighbouring dense bodies are linked by microtubules giving rise to the linear staining patterns in Figure 6. The microtubules extend into the cytosol that contains the nucleus, mitochondria, and other membranes (not shown).

Akhmanova, Microtubule plus-end-tracking proteins: mechanisms and functions. 2005, Pubmed

Akhmanova, Microtubule plus-end-tracking proteins: mechanisms and functions. 2005, Pubmed
Basu, Shaping the actin cytoskeleton using microtubule tips. 2007, Pubmed
Bateman, The Pfam protein families database. 2004, Pubmed
Birukova, Involvement of microtubules and Rho pathway in TGF-beta1-induced lung vascular barrier dysfunction. 2005, Pubmed
Brenner, The genetics of Caenorhabditis elegans. 1974, Pubmed
Caviston, Microtubule motors at the intersection of trafficking and transport. 2006, Pubmed
Chalfie, Organization of neuronal microtubules in the nematode Caenorhabditis elegans. 1979, Pubmed
Chalfie, Developmental genetics of the mechanosensory neurons of Caenorhabditis elegans. 1981, Pubmed
Chalfie, The identification and suppression of inherited neurodegeneration in Caenorhabditis elegans. 1990, Pubmed
Chalfie, Structural and functional diversity in the neuronal microtubules of Caenorhabditis elegans. 1982, Pubmed
Chang, GEF-H1 couples nocodazole-induced microtubule disassembly to cell contractility via RhoA. 2008, Pubmed
Clifford, FOG-2, a novel F-box containing protein, associates with the GLD-1 RNA binding protein and directs male sex determination in the C. elegans hermaphrodite germline. 2000, Pubmed
Cox, Sticky worms: adhesion complexes in C. elegans. 2004, Pubmed
Cueva, Nanoscale organization of the MEC-4 DEG/ENaC sensory mechanotransduction channel in Caenorhabditis elegans touch receptor neurons. 2007, Pubmed
Danowski, Fibroblast contractility and actin organization are stimulated by microtubule inhibitors. 1989, Pubmed
De Keersmaecker, Fusion of EML1 to ABL1 in T-cell acute lymphoblastic leukemia with cryptic t(9;14)(q34;q32). 2005, Pubmed , Echinobase
Dixon, Muscle arm development in Caenorhabditis elegans. 2005, Pubmed
Eichenmuller, The human EMAP-like protein-70 (ELP70) is a microtubule destabilizer that localizes to the mitotic apparatus. 2002, Pubmed , Echinobase
Eichenmüller, Saturable binding of the echinoderm microtubule-associated protein (EMAP) on microtubules, but not filamentous actin or vimentin filaments. 2001, Pubmed , Echinobase
Ernstrom, Genetics of sensory mechanotransduction. 2002, Pubmed
Eudy, Isolation of a novel human homologue of the gene coding for echinoderm microtubule-associated protein (EMAP) from the Usher syndrome type 1a locus at 14q32. 1997, Pubmed , Echinobase
Francis, Muscle cell attachment in Caenorhabditis elegans. 1991, Pubmed
Francis, Muscle organization in Caenorhabditis elegans: localization of proteins implicated in thin filament attachment and I-band organization. 1985, Pubmed
Hall, Kinesin-related gene unc-104 is required for axonal transport of synaptic vesicles in C. elegans. 1991, Pubmed
Hall, Rho GTPases and the actin cytoskeleton. 1998, Pubmed
Hamill, Purification of a WD repeat protein, EMAP, that promotes microtubule dynamics through an inhibition of rescue. 1998, Pubmed , Echinobase
Hedgecock, The unc-5, unc-6, and unc-40 genes guide circumferential migrations of pioneer axons and mesodermal cells on the epidermis in C. elegans. 1990, Pubmed
Heidebrecht, Cloning and localization of C2orf2(ropp120), a previously unknown WD repeat protein. 2000, Pubmed
Houtman, Echinoderm microtubule-associated protein like protein 4, a member of the echinoderm microtubule-associated protein family, stabilizes microtubules. 2007, Pubmed , Echinobase
Howard, Microtubule polymerases and depolymerases. 2007, Pubmed
Hresko, Myotactin, a novel hypodermal protein involved in muscle-cell adhesion in Caenorhabditis elegans. 1999, Pubmed
Inamura, EML4-ALK fusion is linked to histological characteristics in a subset of lung cancers. 2008, Pubmed , Echinobase
Kolodney, Contraction due to microtubule disruption is associated with increased phosphorylation of myosin regulatory light chain. 1995, Pubmed
Krendel, Nucleotide exchange factor GEF-H1 mediates cross-talk between microtubules and the actin cytoskeleton. 2002, Pubmed
Kwok, Microtubule flux: drivers wanted. 2007, Pubmed
Labouesse, Epithelial junctions and attachments. 2006, Pubmed
Lepley, Sequence and expression patterns of a human EMAP-related protein-2 (HuEMAP-2). 1999, Pubmed , Echinobase
Li, Molecular characterization of the 77-kDa echinoderm microtubule-associated protein. Homology to the beta-transducin family. 1994, Pubmed , Echinobase
Li, The 77-kDa echinoderm microtubule-associated protein (EMAP) shares epitopes with the mammalian brain MAPs, MAP-2 and tau. 1998, Pubmed , Echinobase
Matthews, ZYG-9, a Caenorhabditis elegans protein required for microtubule organization and function, is a component of meiotic and mitotic spindle poles. 1998, Pubmed
Miller, Immunofluorescence microscopy. 1995, Pubmed
Moerman, Sarcomere assembly in C. elegans muscle. 2006, Pubmed
O'Connor, Eml5, a novel WD40 domain protein expressed in rat brain. 2004, Pubmed , Echinobase
Perkins, Mutant sensory cilia in the nematode Caenorhabditis elegans. 1986, Pubmed
Picking, Cloning, expression, and affinity purification of recombinant Shigella flexneri invasion plasmid antigens IpaB and IpaC. 1996, Pubmed
Pollmann, Human EML4, a novel member of the EMAP family, is essential for microtubule formation. 2006, Pubmed
Siegrist, Microtubule-induced cortical cell polarity. 2007, Pubmed
Small, How do microtubules guide migrating cells? 2002, Pubmed
Soda, Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer. 2007, Pubmed , Echinobase
Sulston, The Caenorhabditis elegans male: postembryonic development of nongonadal structures. 1980, Pubmed
Suprenant, Conservation of the WD-repeat, microtubule-binding protein, EMAP, in sea urchins, humans, and the nematode C. elegans. 2000, Pubmed , Echinobase
Suprenant, Sea urchin microtubules. 1995, Pubmed , Echinobase
Tegha-Dunghu, EML3 is a nuclear microtubule-binding protein required for the correct alignment of chromosomes in metaphase. 2008, Pubmed
White, The structure of the nervous system of the nematode Caenorhabditis elegans. 1986, Pubmed