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Phosphorylation mediates the nuclear targeting of the maize Rab17 protein
Author(s) -
Jensen Anders B.,
Goday Adela,
Figueras Mercè,
Jessop Alison C.,
Pagès Montserrat
Publication year - 1998
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1998.00069.x
Subject(s) - nuclear localization sequence , biology , arabidopsis thaliana , nuclear transport , phosphorylation , arabidopsis , cytoplasm , fusion protein , amino acid , transgene , kinase , nls , mutation , microbiology and biotechnology , biochemistry , cell nucleus , mutant , gene , recombinant dna
Summary The maize abscisic acid‐responsive Rab17 protein localizes to the nucleus and cytoplasm in maize cells. In‐frame fusion of Rab17 to the reporter protein β‐glucuronidase (GUS) directed GUS to the nucleus and cytoplasm in transgenic Arabidopsis thaliana and in transiently transformed onion cells. Analysis of chimeric constructs identified one region between amino acid positions 66–96, which was necessary for targeting GUS to the nucleus. This region contains a serine cluster followed by a putative consensus site for protein kinase CK2 phosphorylation, and a stretch of basic amino acids resembling the simian virus 40 large T antigen‐type nuclear localization signal (NLS). Mutation of two basic amino acids in the putative NLS had a weak effect on nuclear targeting in the onion cell system and did not modify the percentage of nuclear fusion protein in the Arabidopsis cells. The mutation of three amino acids in the consensus site for CK2 recognition resulted in the absence of in vitro phosphorylated forms of Rab17 and in a strong decrease of GUS enzymatic activity in isolated nuclei of transgenic Arabidopsis. These results suggest that phosphorylation of Rab17 by protein kinase CK2 is the relevant step for its nuclear location, either by facilitating binding to specific proteins or as a direct part of the nuclear targeting apparatus.