The gene for stinging nettle lectin (Urtica dioica agglutinin) encodes both a lectin and a chitinase.
Chitin-binding proteins are present in a wide range of plant species, including both monocots and dicots, even though these plants contain no chitin. To investigate the relationship between in vitro antifungal and insecticidal activities of chitin-binding proteins and their unknown endogenous functions, the stinging nettle lectin (Urtica dioica agglutinin, UDA) cDNA was cloned using a synthetic gene as the probe. The nettle lectin cDNA clone contained an open reading frame encoding 374 amino acids. Analysis of the deduced amino acid sequence revealed a 21-amino acid putative signal sequence and the 86 amino acids encoding the two chitin-binding domains of nettle lectin. These domains were fused to a 19-amino acid "spacer" domain and a 244-amino acid carboxyl extension with partial identity to a chitinase catalytic domain. The authenticity of the cDNA clone was confirmed by deduced amino acid sequence identity with sequence data obtained from tryptic digests, RNA gel blot, and polymerase chain reaction analyses. RNA gel blot analysis also showed the nettle lectin message was present primarily in rhizomes and inflorescence (with immature seeds) but not in leaves or stems. Chitinase enzymatic activity was found when the chitinase-like domain alone or the chitinase-like domain with the chitin-binding domains were expressed in Escherichia coli. This is the first example of a chitin-binding protein with both a duplication of the 43-amino acid chitin-binding domain and a fusion of the chitin-binding domains to a structurally unrelated domain, the chitinase domain.
Lerner
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DR
Department of Energy Plant Research Laboratory, Michigan State University, East Lansing 48824-1312.
Raikhel
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NV
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D037102:Lectins / Q000235:genetics*; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010947:Plants, Toxic / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000235:genetics; D012313:RNA / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D037102:Lectins; D037121:Plant Lectins; D021382:Protein Sorting Signals; D012313:RNA; D004247:DNA; D002688:Chitinases