Nucleic Acids Research, Vol 27, Issue 24 4703-4709, Copyright © 1999 by Oxford University Press
Y Le, S Gagneten, D Tombaccini, B Bethke and B Sauer
The Cre DNA recombinase of bacteriophage P1 has become a useful tool for
genomic manipulation in mice and other eukaryotes. Because Cre is of
prokaryotic origin, the 38 kDa protein has been presumed to gain access to
the eukaryotic nucleus simply because it is sufficiently small to pass
through the nuclear pore by passive diffusion. Instead, we show here that
Cre carries nuclear targeting determinants that efficiently direct Cre
entry into the nucleus of mammalian cells. Fusions of Cre with green
fluorescent protein (GFP) identified two regions that are necessary for
nuclear localization. Region I contains a cluster of basic amino acids that
is essential for nuclear localization and which resembles a bipartite-like
nuclear localization signal. Region II exhibits a beta-sheet structure with
which the bipartite motif may interact. However, neither region is by
itself sufficient for nuclear localization. Nuclear transport in vitro with
a 98 kDa GFP-Cre fusion protein shows that Cre does not gain access to the
nucleus by passive diffusion, but instead enters the nucleus by means of an
energy-dependent process. Thus, Cre is one of the few prokaryotic proteins
that have been shown to carry determinants that allow it to target the
eukaryotic nucleus.
ARTICLES
Nuclear targeting determinants of the phage P1 cre DNA recombinase
Developmental Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA.
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