Complex formation between Tap and p15 affects binding to FG-repeat nucleoporins and nucleocytoplasmic shuttling

J Biol Chem. 2002 Mar 15;277(11):9242-6. doi: 10.1074/jbc.M110007200. Epub 2001 Dec 27.

Abstract

Mammalian Tap-p15 and yeast Mex67p-Mtr2p are conserved and essential mRNA export factor complexes that transport mRNPs through the nuclear pore. Here, we report that the small subunit p15 affects the binding of the large subunit Tap to repeat nucleoporins. BIAcore measurements revealed that recombinant Tap binds with high affinity (K(d) in the nm range) to repeat nucleoporins and dissociates from them very slowly. In contrast, when recombinant Tap was bound to p15, the derived heterodimeric complex exhibited a significant lower affinity to FG-repeat nucleoporins (K(d) in the microm range). Furthermore, when recombinant Tap lacking the N-terminal nuclear localization sequences (TapDeltaNLS) was microinjected in mammalian cells, it did not shuttle; however, TapDeltaNLS with bound p15 efficiently shuttles between nucleus and cytoplasm. We conclude that heterodimerization of Tap and p15 is required for shuttling of the functional Tap-p15 mRNA exporter complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 2
  • ATP-Binding Cassette Transporters
  • Animals
  • Carrier Proteins / metabolism*
  • Cattle
  • Cell Cycle Proteins*
  • Cell Nucleus / metabolism*
  • Cyclin-Dependent Kinase Inhibitor p15
  • Cytoplasm / metabolism*
  • Dimerization
  • HeLa Cells
  • Humans
  • Nuclear Pore Complex Proteins / chemistry
  • Nuclear Pore Complex Proteins / metabolism*
  • Nucleocytoplasmic Transport Proteins*
  • Repetitive Sequences, Amino Acid
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*
  • Tumor Suppressor Proteins*

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 2
  • ATP-Binding Cassette Transporters
  • CDKN2B protein, human
  • Carrier Proteins
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p15
  • NXT1 protein, human
  • Nuclear Pore Complex Proteins
  • Nucleocytoplasmic Transport Proteins
  • TAP1 protein, human
  • Transcription Factors
  • Tumor Suppressor Proteins