Binding of recombinant but not endogenous prion protein to DNA Causes DNA internalization and expression in mammalian cells

Shaoman Yin, Xingjun Fan, Shuiliang Yu, Chaoyang Li, Man Sun Sy

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Recombinant prion protein, rPrP, binds DNA. Both the KKRPK motif and the octapeptide repeat region of rPrP are essential for maximal binding. rPrP with pathogenic insertional mutations binds more DNA than wild-type rPrP. DNA promotes the aggregation of rPrP and protects its N terminus from proteinase K digestion. When rPrP is mixed with an expression plasmid and Ca2+, the rPrP·DNA complex is taken up by mammalian cells leading to gene expression. In the presence of Ca2+, rPrP by itself is also taken up by cells in a temperature- and pinocytosis-dependent manner. Cells do not take up rPrPΔKKRPK, which lacks the KKRPK motif. Thus, rPrP is the carrier for DNA and the KKRPK motif is essential for its uptake. When mixed with DNA, a pentapeptide KKRPK, but not KKKKK, is sufficient for DNA internalization and expression. In contrast, whereas the normal cellular prion protein, PrPC, on the cell surface can also internalize DNA, the imported DNA is not expressed. These findings may have relevance to the normal functions of PrPC and the pathogenic mechanisms of human prion disease.

Original languageEnglish (US)
Pages (from-to)25446-25454
Number of pages9
JournalJournal of Biological Chemistry
Volume283
Issue number37
DOIs
StatePublished - Sep 12 2008

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Cells
DNA
PrPC Proteins
Pinocytosis
Endopeptidase K
Nucleotide Motifs
Prion Diseases
Prions
Prion Proteins
Recombinant Proteins
Digestion
Gene expression
Plasmids
Agglomeration
Gene Expression
Mutation
Temperature

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Binding of recombinant but not endogenous prion protein to DNA Causes DNA internalization and expression in mammalian cells. / Yin, Shaoman; Fan, Xingjun; Yu, Shuiliang; Li, Chaoyang; Sy, Man Sun.

In: Journal of Biological Chemistry, Vol. 283, No. 37, 12.09.2008, p. 25446-25454.

Research output: Contribution to journalArticle

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AU - Sy, Man Sun

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