Structural and functional characterization of a novel, host penetration-related pectate lyase from the potato cyst nematode Globodera rostochiensis

U. Kudla, A. Milac, Qin Ling, H.A. Overmars, E.H.A. Roze, M.H.M. Holterman, A.J. Petrescu, A. Goverse, J. Bakker, J. Helder, G. Smant

Research output: Contribution to journalArticleAcademicpeer-review

30 Citations (Scopus)

Abstract

The cell wall, a strong extraprotoplasmic layer surrounding plant cells that mainly consists of a variety of polysaccharides, constitutes a major barrier for potential parasites. Plant-parasitic nematodes are well equipped to overcome this barrier as they produce and secrete cell-wall-degrading enzymes. Expression profiling of various life stages of the potato cyst nematode Globodera rostochiensis revealed a novel pectate lyase gene (Gr-pel2, 759 bp). The Gr-PEL2 protein showed highest similarity to pectate lyases from the facultative plant-parasitic nematodes Bursaphelenchus mucronatus and B. xylophilus and the soil-inhabiting saprophytic Streptomyces and Frankia species (i.e. 40-42% identity and 58-60% similarity), whereas only a remote relatedness to the previously identified Gr-PEL1 was observed (i.e. 28% identity and 43% similarity). Transient expression of Gr-pel2 in leaves of Nicotiana benthamiana resulted in severe malformations of the infiltrated tissues, not relating to maceration and soft rot symptoms. Ca2+ is known to be essential for pectate lyase activity, and the most likely calcium-binding site was identified in the Gr-PEL2 protein by combining homology modelling of the three-dimensional structure, site-directed mutagenesis and transient expression in leaves. A highly charged cleft in Gr-PEL2, which is likely to be involved in substrate binding and which is also significantly more hydrophobic in Gr-PEL1, was shown to be essential for protein activity. Our results underline the broad spectrum of pectate lyases and cell-wall-degrading enzymes necessary for successful parasitism by cyst nematodes
Original languageEnglish
Pages (from-to)293-305
JournalMolecular Plant Pathology
Volume8
Issue number3
DOIs
Publication statusPublished - 2007

Keywords

  • subventral esophageal glands
  • protein secondary structure
  • heterodera-glycines
  • developmental expression
  • erwinia-chrysanthemi
  • structure prediction
  • new-generation
  • agrobacterium
  • identification
  • transformation

Fingerprint

Dive into the research topics of 'Structural and functional characterization of a novel, host penetration-related pectate lyase from the potato cyst nematode Globodera rostochiensis'. Together they form a unique fingerprint.

Cite this