Faculty Opinions recommendation of Nuclear localization signal receptor affinity correlates with in vivo localization in Saccharomyces cerevisiae.

Initiation of eukaryotic genome duplication begins when a six-subunit origin recognition complex (ORC) binds to DNA. However, the mechanism by which this occurs in vivo and the roles played by individual subunits appear to differ significantly among organisms. Previous studies identified a soluble human ORC(2–5) complex in the nucleus, an ORC(1–5) complex bound to chromatin, and an Orc6 protein that binds weakly, if at all, to other ORC subunits. Here we show that stable ORC(1–6) complexes also can be purified from human cell extracts and that Orc6 and Orc1 each contain a single nuclear localization signal that is essential for nuclear localization but not for ORC assembly. The Orc6 nuclear localization signal, which is essential for Orc6 function, is facilitated by phosphorylation at its cyclin-dependent kinase consensus site and by association with Kpna6/1, nuclear transport proteins that did not co-purify with other ORC subunits. These and other results support a model in which Orc6, Orc1, and ORC(2–5) are transported independently to the nucleus where they can either assemble into ORC(1–6) or function individually.

Download Full-text

Analysis of a predicted nuclear localization signal: implications for the intracellular localization and function of the Saccharomyces cerevisiae RNA-binding protein Scp160

Nucleic Acids Research ◽

10.1093/nar/gkm776 ◽

2007 ◽

Vol 35 (20) ◽

pp. 6862-6869 ◽

Cited By ~ 5

Author(s):

M. A. Brykailo ◽

L. M. McLane ◽

J. Fridovich-Keil ◽

A. H. Corbett

Keyword(s):

Saccharomyces Cerevisiae ◽

Nuclear Localization ◽

Nuclear Localization Signal ◽

Rna Binding ◽

Binding Protein ◽

Intracellular Localization ◽

Rna Binding Protein ◽

Localization Signal ◽

And Function

Download Full-text

The Nuclear Localization Signal of the Prrs Virus Nucleocapsid Protein Modulates Viral Replication in vitro and Antibody Response in vivo

Advances in Experimental Medicine and Biology - The Nidoviruses ◽

10.1007/978-0-387-33012-9_25 ◽

2006 ◽

pp. 145-148 ◽

Cited By ~ 10

Author(s):

Changhee Lee ◽

Douglas C. Hodgins ◽

Jay G. Calvert ◽

Siao-Kun Wan Welch ◽

Rika Jolie ◽

...

Keyword(s):

Antibody Response ◽

Viral Replication ◽

Nuclear Localization ◽

Nuclear Localization Signal ◽

Nucleocapsid Protein ◽

Prrs Virus ◽

Localization Signal

Download Full-text

Degradation of Saccharomyces cerevisiae Transcription Factor Gcn4 Requires a C-Terminal Nuclear Localization Signal in the Cyclin Pcl5

Eukaryotic Cell ◽

10.1128/ec.00324-08 ◽

2009 ◽

Vol 8 (4) ◽

pp. 496-510 ◽

Cited By ~ 8

Author(s):

Katrin Streckfuss-Bömeke ◽

Florian Schulze ◽

Britta Herzog ◽

Eva Scholz ◽

Gerhard H. Braus

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acids ◽

Nuclear Localization ◽

Nuclear Localization Signal ◽

Ubiquitin Proteasome System ◽

Cyclin Dependent Kinase ◽

General Amino Acid Control ◽

Nuclear Domain ◽

Ubiquitin Proteasome ◽

Localization Signal

ABSTRACT Pcl5 is a Saccharomyces cerevisiae cyclin that directs the phosphorylation of the general amino acid control transcriptional activator Gcn4 by the cyclin-dependent kinase (CDK) Pho85. Phosphorylation of Gcn4 by Pho85/Pcl5 initiates its degradation via the ubiquitin/proteasome system and is regulated by the availability of amino acids. In this study, we show that Pcl5 is a nuclear protein and that artificial dislocation of Pcl5 into the cytoplasm prevents the degradation of Gcn4. Nuclear localization of Pcl5 depends on the β-importin Kap95 and does not require Pho85, Gcn4, or the CDK inhibitor Pho81. Pcl5 nuclear import is independent on the availability of amino acids and is mediated by sequences in its C-terminal domain. The nuclear localization signal is distinct from other functional domains of Pcl5. This is corroborated by a C-terminally truncated Pcl5 variant, which carries the N-terminal nuclear domain of Pho80. This hybrid is still able to fulfill Pcl5 function, whereas Pho80, which is another Pho85 interacting cyclin, does not mediate Gcn4 degradation.

Download Full-text

Invading the Yeast Nucleus: a Nuclear Localization Signal at the C Terminus of Ty1 Integrase Is Required for Transposition In Vivo

Molecular and Cellular Biology ◽

10.1128/mcb.18.2.1115 ◽

1998 ◽

Vol 18 (2) ◽

pp. 1115-1124 ◽

Cited By ~ 63

Author(s):

Margaret A. Kenna ◽

Carrie Baker Brachmann ◽

Scott E. Devine ◽

Jef D. Boeke

Keyword(s):

Nuclear Localization ◽

Nuclear Localization Signal ◽

Reverse Transcriptase Activity ◽

Deletion Mutants ◽

Amino Acid Residues ◽

Wild Type ◽

C Terminus ◽

Localization Signal

ABSTRACT Retrotransposon Ty1 faces a formidable cell barrier during transposition—the yeast nuclear membrane which remains intact throughout the cell cycle. We investigated the mechanism by which transposition intermediates are transported from the cytoplasm (the presumed site of Ty1 DNA synthesis) to the nucleus, where they are integrated into the genome. Ty1 integrase has a nuclear localization signal (NLS) at its C terminus. Both full-length integrase and a C-terminal fragment localize to the nucleus. C-terminal deletion mutants in Ty1 integrase were used to map the putative NLS to the last 74 amino acid residues of integrase. Mutations in basic segments within this region decreased retrotransposition at least 50-fold in vivo. Furthermore, these mutant integrase proteins failed to localize to the nucleus. Production of virus-like particles, reverse transcriptase activity, and complete in vitro Ty1 integration resembled wild-type levels, consistent with failure of the mutant integrases to enter the nucleus.

Download Full-text