NtSCP1 from tobacco is an extracellular serine carboxypeptidase III that has an impact on cell elongation

Plant Physiol. 2012 Mar;158(3):1220-9. doi: 10.1104/pp.111.192088. Epub 2012 Jan 3.

Abstract

The leaf extracellular space contains several peptidases, most of which are of unknown function. We isolated cDNAs for two extracellular serine carboxypeptidase III genes from tobacco (Nicotiana tabacum), NtSCP1 and NtSCP2, belonging to a phylogenetic clade not yet functionally characterized in plants. NtSCP1 and NtSCP2 are orthologs derived from the two ancestors of tobacco. Reverse transcription-polymerase chain reaction analysis showed that NtSCP1 and NtSCP2 are expressed in root, stem, leaf, and flower tissues. Expression analysis of the β-glucuronidase reporter gene fused to the NtSCP1 transcription promoter region confirmed this expression profile. Western blotting of NtSCP1 and expression of an NtSCP1-green fluorescent protein fusion protein showed that the protein is located in the extracellular space of tobacco leaves and culture cells. Purified His-tagged NtSCP1 had carboxypeptidase activity in vitro. Transgenic tobacco plants overexpressing NtSCP1 showed a reduced flower length due to a decrease in cell size. Etiolated seedlings of these transgenic plants had shorter hypocotyls. These data provide support for a role of an extracellular type III carboxypeptidase in the control of cell elongation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carboxypeptidases / genetics
  • Carboxypeptidases / metabolism*
  • Cell Enlargement*
  • Cloning, Molecular
  • Culture Media / metabolism
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • DNA, Plant / genetics
  • DNA, Plant / metabolism*
  • Enzyme Activation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Extracellular Space / genetics
  • Extracellular Space / metabolism
  • Gene Expression Regulation, Plant
  • Genes, Reporter
  • Green Fluorescent Proteins / metabolism
  • Molecular Sequence Data
  • Nicotiana / enzymology*
  • Nicotiana / genetics
  • Nicotiana / growth & development
  • Phylogeny
  • Plant Components, Aerial / genetics
  • Plant Components, Aerial / growth & development
  • Plant Components, Aerial / metabolism
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plant Roots / genetics
  • Plant Roots / growth & development
  • Plant Roots / metabolism
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / growth & development
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Culture Media
  • DNA, Complementary
  • DNA, Plant
  • Plant Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Carboxypeptidases
  • serine carboxypeptidase

Associated data

  • GENBANK/ABQ65859
  • GENBANK/ABQ65860
  • GENBANK/GU734644