Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein

C. Spencer Yost, Charles Lopez, Stanley B. Prusiner, Richard M. Myers, Vishwanath R. Lingappa

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

A UNIVERSAL feature of integral transmembrane proteins is a hydrophobic peptide segment that spans the lipid bilayer. These hydrophobic domains are important for terminating the translocation of the polypeptide chain across the membrane of the endoplasmic reticulum (a process termed stop transfer) and for integrating the protein into the bilayer1-5. But a role for extracytoplasmic sequences in stop transfer and transmembrane integration has not previously been shown. Recently, a sequence which directs an unusual mode of stop transfer has been identified in the prion protein. This brain glycoprotein exists in two isoforms6-8, which are identical both in primary amino-acid sequence and in containing phosphatidylinositol glycolipid linkages at their C termini, which can be cleaved by a phophatidylinositol-specific phospholipase C9. But only one of the isoforms (PrPC) is released from cells on treatment with this phospholipase10, indicating that the two isoforms have either different subcellular locations or transmembrane orientations. Consistent with this is the observation of two different topological forms in cell-free systems11,12. An unusual topogenic sequence in the prion protein seems to direct these alternative topologies (manuscript in preparation). In the wheat-germ translation system, this sequence directs nascent chains to a transmembrane orientation; by contrast, in the rabbit reticulocyte lysate system, this sequence fails to cause stop transfer of most nascent chains. We have now investigated determinants in this unusual topogenic sequence that direct transmembrane topology, and have demonstrated that (1) a lumenally disposed charged domain is required for stop transfer at the adjacent hydrophobic domain, (2) a precise spatial relationship between these domains is essential for efficient stop transfer, and (3) codons encompassing this hydrophilic extracytoplasmic domain confer transmembrane topology to a heterologous protein when engineered adjacent to the codons for a normally translocated hydrophobic domain. These results identify an unexpected functional domain for stop transfer in the prion protein and have implications for the mechanism of membrane protein biogenesis.

Original languageEnglish (US)
Pages (from-to)669-672
Number of pages4
JournalNature
Volume343
Issue number6259
StatePublished - Feb 15 1990
Externally publishedYes

Fingerprint

Codon
Protein Isoforms
Peptides
Proteins
Manuscripts
Phospholipases
Reticulocytes
Glycolipids
Lipid Bilayers
Phosphatidylinositols
Endoplasmic Reticulum
Triticum
Amino Acid Sequence
Glycoproteins
Membrane Proteins
Observation
Rabbits
Membranes
Brain
Prion Proteins

ASJC Scopus subject areas

  • General

Cite this

Yost, C. S., Lopez, C., Prusiner, S. B., Myers, R. M., & Lingappa, V. R. (1990). Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein. Nature, 343(6259), 669-672.

Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein. / Yost, C. Spencer; Lopez, Charles; Prusiner, Stanley B.; Myers, Richard M.; Lingappa, Vishwanath R.

In: Nature, Vol. 343, No. 6259, 15.02.1990, p. 669-672.

Research output: Contribution to journalArticle

Yost, CS, Lopez, C, Prusiner, SB, Myers, RM & Lingappa, VR 1990, 'Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein', Nature, vol. 343, no. 6259, pp. 669-672.
Yost CS, Lopez C, Prusiner SB, Myers RM, Lingappa VR. Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein. Nature. 1990 Feb 15;343(6259):669-672.
Yost, C. Spencer ; Lopez, Charles ; Prusiner, Stanley B. ; Myers, Richard M. ; Lingappa, Vishwanath R. / Non-hydrophobic extracytoplasmic determinant of stop transfer in the prion protein. In: Nature. 1990 ; Vol. 343, No. 6259. pp. 669-672.
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