Identification of functional PDZ domain binding sites in several human proteins

Mol Biol Rep. 2000;27(4):217-24. doi: 10.1023/a:1011008313677.

Abstract

TIP-15 was previously identified as a cellular protein that can bind to the C-terminal end of the HTLV-1 Tax protein via its two PDZ domains. The sequence of the N-terminal part of TIP-15 is identical to that of the synaptic protein PSD-95. Both proteins are likely to be produced from the same gene by alternative splicing. Whereas expression of the PSD-95 mRNA was detected only with brain RNAs, that of TIP-15 was detected with RNAs from thymus, brain, skeletal muscle and Jurkat cells. The TIP-15 protein exhibits an apparent molecular weight of 40 kD and is weakly expressed in T cell lines. A two-hybrid screen performed with TIP-15 as bait revealed the presence of a PDZ binding site (PDZ-BS) in the following proteins: Lysyl tRNA synthetase, 6-phosphogluconolactonase (6-GPL), Stress-activated protein kinase 3 (SAPK3), NET-1, Diacylglycerol kinase zeta, MTMR1, MCM7, and hSec8. The sequence at the C-terminal ends of these proteins matches the X-S/T-X-V-COOH consensus previously defined for PDZ-BSs, with the exception of 6-GPL and SAPK3 which include a leucine as the C-terminal residue. For Lysyl tRNA synthetase, NET1, MTMR1 and hSec8, binding to TIP-15 was confirmed by co-immunoprecipitation experiments performed with the extracts of transfected COS7 cells. These results show the existence of functional PDZ-BSs in these proteins, but future studies will be necessary to establish whether or not TIP-15 represents a physiological partner. The significance of the presence of a PDZ-BS in these various proteins is discussed with respect to their function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Brain / metabolism
  • COS Cells
  • Carboxylic Ester Hydrolases / chemistry
  • Carrier Proteins / chemistry
  • Cell Cycle Proteins / chemistry
  • DNA-Binding Proteins / chemistry
  • Diacylglycerol Kinase / chemistry
  • Disks Large Homolog 4 Protein
  • Gene Products, tax / chemistry*
  • Guanylate Kinases
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Jurkat Cells
  • Membrane Proteins
  • Mice
  • Minichromosome Maintenance Complex Component 7
  • Mitogen-Activated Protein Kinase 12
  • Mitogen-Activated Protein Kinases / chemistry
  • Molecular Sequence Data
  • Muscle, Skeletal / metabolism
  • Nerve Tissue Proteins / metabolism
  • Nuclear Proteins / chemistry
  • Oncogene Proteins / chemistry
  • Plasmids / metabolism
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary*
  • Protein Tyrosine Phosphatases / chemistry
  • Protein Tyrosine Phosphatases, Non-Receptor
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Synapses / metabolism
  • Two-Hybrid System Techniques
  • Vesicular Transport Proteins
  • beta-Galactosidase / metabolism

Substances

  • Carrier Proteins
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Disks Large Homolog 4 Protein
  • Dlg4 protein, mouse
  • EXOC4 protein, human
  • Gene Products, tax
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • NET1 protein, human
  • Nerve Tissue Proteins
  • Net1 protein, mouse
  • Nuclear Proteins
  • Oncogene Proteins
  • RNA, Messenger
  • Sec8l1 protein, mouse
  • Vesicular Transport Proteins
  • postsynaptic density proteins
  • Mitogen-Activated Protein Kinase 12
  • Diacylglycerol Kinase
  • Mitogen-Activated Protein Kinases
  • Guanylate Kinases
  • Carboxylic Ester Hydrolases
  • 6-phosphogluconolactonase
  • MTMR1 protein, human
  • Protein Tyrosine Phosphatases
  • Protein Tyrosine Phosphatases, Non-Receptor
  • beta-Galactosidase
  • MCM7 protein, human
  • Mcm7 protein, mouse
  • Minichromosome Maintenance Complex Component 7