Identification, localization, and functional implications of an abundant nematode annexin.
Cultures of the nematode C. elegans were examined for the presence of calcium-dependent, phospholipid-binding proteins of the annexin class. A single protein of apparent mass on SDS-polyacrylamide gels of 32 kD was isolated from soluble extracts of nematode cultures on the basis of its ability to bind to phospholipids in a calcium-dependent manner. After verification of the protein as an annexin by peptide sequencing, an antiserum to the protein was prepared and used to isolate a corresponding cDNA from an expression library in phage lambda gt11. The encoded protein, herein referred to as the nex-1 annexin, has a mass of 35 kD and is 36-42% identical in sequence to 10 known mammalian annexins. Several unique modifications were found in the portions of the sequence corresponding to calcium-binding sites. Possible phosphorylation sites in the NH2-terminal domain of the nematode annexin correspond to those of mammalian annexins. The gene for this annexin (nex-1) was physically mapped to chromosome III in the vicinity of the dpy-17 genetic marker. Two other annexin genes (nex-2 and nex-3) were also identified in chromosome III sequences reported by the nematode genomic sequencing project (Sulston, J., Z. Du, K. Thomas, R. Wilson, L. Hillier, R. Staden, N. Halloran, P. Green, J. Thierry-Mieg, L. Qiu, et al. 1992. Nature (Lond.). 356:37-41). The nex-1 annexin was localized in the nematode by immunofluorescence and by electron microscopy using immunogold labeling. The protein is associated with membrane systems of the secretory gland cells of the pharynx, with sites of cuticle formation in the grinder in the pharynx, with yolk granules in oocytes, with the uterine wall and vulva, and with membrane systems in the spermathecal valve. The presence of the annexin in association with the membranes of the spermathecal valve suggests a novel function of the protein in the folding and unfolding of these membranes as eggs pass through the valve. The localizations also indicate roles for the annexin corresponding to those proposed in mammalian systems in membrane trafficking, collagen deposition, and extracellular matrix formation.
Creutz
C E
CE
Department of Pharmacology, University of Virginia, Charlottesville 22908, USA.
Snyder
S L
SL
Daigle
S N
SN
Redick
J
J
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000737:chemistry* / Q000648:ultrastructure; D029742:Caenorhabditis elegans Proteins; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002874:Chromosome Mapping; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D017238:Genes, Helminth; D005836:Genitalia, Female / Q000737:chemistry / Q000648:ultrastructure; D015801:Helminth Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D013045:Species Specificity
D017302:Annexins; D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary; D015801:Helminth Proteins; C098890:Nex-1 protein, C elegans; D011994:Recombinant Proteins