Cloning of a Tobacco Apoplasmic Invertase Inhibitor1
- 1 February 1998
- journal article
- Published by Oxford University Press (OUP) in Plant Physiology
- Vol. 116 (2) , 733-742
- https://doi.org/10.1104/pp.116.2.733
Abstract
Higher plants express several isoforms of vacuolar and cell wall invertases (CWI), some of which are inactivated by inhibitory proteins at certain stages of plant development. We have purified an apoplasmic inhibitor (INH) of tobacco (Nicotiana tabacum) CWI to homogeneity. Based on sequences from tryptic fragments, we have isolated a full-length INH-encoding cDNA clone (Nt-inh1) via a reverse transcriptase-polymerase chain reaction. Southern-blot analysis revealed that INH is encoded by a single- or low-copy gene. Comparison with expressed sequence tag clones from Arabidopsis thaliana and Citrus unshiu indicated the presence of Nt-inh1-related proteins in other plants. The recombinantNt-inh1-encoded protein inhibits CWI from tobacco andChenopodium rubrum suspension-cultured cells and vacuolar invertase from tomato (Lycopersicon esculentum) fruit, whereas yeast invertase is not affected. However, only in the homologous system is the inhibition modulated by the concentration of Suc as previously shown for INH isolated from tobacco cells. Highly specific binding of INH to CWI could be shown by affinity chromatography of a total cell wall protein fraction on immobilized recombinant Nt-inh1 protein. RNA-blot analysis of relative transcript ratios for Nt-inh1 and CWI in different parts of adult tobacco plants revealed that the expression of both proteins is not always coordinate.Keywords
This publication has 38 references indexed in Scilit:
- Co‐ordinated induction of mRNAs for extracellular invertase and a glucose transporter in Chenopodium rubrum by cytokininsThe Plant Journal, 1997
- The sink-specific and stress-regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors, and pathogen challenge.Plant Cell, 1996
- The Miniature1 Seed Locus of Maize Encodes a Cell Wall Invertase Required for Normal Development of Endosperm and Maternal Cells in the PedicelPlant Cell, 1996
- Sucrose protects cell wall invertase but not vacuolar invertase against proteinaceous inhibitorsFEBS Letters, 1996
- A Tobacco cDNA Coding for Cell-Wall InvertasePlant Physiology, 1995
- A sink‐specific H+/monosaccharide co‐transporter from Nicotiana tabacum: cloning and heterologous expression in baker’s yeastThe Plant Journal, 1993
- The Maize Invertase-Deficient miniature-1 Seed Mutation Is Associated with Aberrant Pedicel and Endosperm Development.Plant Cell, 1992
- The Maize Invertase-Deficient miniature-1 Seed Mutation Is Associated with Aberrant Pedicel and Endosperm DevelopmentPlant Cell, 1992
- Physical properties of the cell wall of photoautotrophic suspension cells fromChenopodium rubrum L.Planta, 1988
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979