Karyopherin? proteins mediate nucleocytoplasmic transport through recognition of distinct nuclear localization or export signals (NLSs or NESs). Although there are 19 known Kaps in humans, only three classes of signals for Kap?/Kap?1, Kap?2 (or Transportin) and CRM1, respectively, are known at this time. Most other Kaps recognize diverse sequences that occlude the identity of their signals. This proposal describes structural and biochemical analyses of import systems Kap?2 and Imp5/Kap121 as well as the export-Kap CRM1. Kap?2 mediates nuclear import of RNA binding proteins through their PY-NLSs. Our discovery and understanding of the PY-NLS contributed to the discovery of a defective PY-NLS that causes a subset of amyotrophic lateral sclerosis (ALS). Here, we propose to understand the role of Kap?2 in protein mislocalization and aggregation in ALS. In another aim, we will apply the combined structural, biochemical and bioinformatics approach that we used to discover the PY-NLS to the Imp5/Kap121p pathway with the goal of discovering a set of characteristics to unify the diverse sequences recognized by the Kaps, hence revealing a new class of NLS. Finally, we will study CRM1, which mediates nuclear export of hundreds of proteins;most of them identified using the small molecule inhibitor Leptomycin B. This proposal aims to determine the mechanism of how CRM1 mediates conjugation of leptomycin B to inhibit nuclear export.

Public Health Relevance

Many proteins are mislocalized in diseases such as amyotrophic lateral sclerosis and cancer. It is important to understand how proteins are targeted correctly since correcting protein mislocalization may be a valid disease therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM069909-10
Application #
8440064
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Ainsztein, Alexandra M
Project Start
2004-02-01
Project End
2017-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
10
Fiscal Year
2013
Total Cost
$333,488
Indirect Cost
$123,747
Name
University of Texas Sw Medical Center Dallas
Department
Pharmacology
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
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Soniat, Michael; CaÄŸatay, Tolga; Chook, Yuh Min (2016) Recognition Elements in the Histone H3 and H4 Tails for Seven Different Importins. J Biol Chem 291:21171-21183
Hing, Z A; Fung, H Y J; Ranganathan, P et al. (2016) Next-generation XPO1 inhibitor shows improved efficacy and in vivo tolerability in hematological malignancies. Leukemia 30:2364-2372
Soniat, Michael; Chook, Yuh Min (2016) Karyopherin-β2 Recognition of a PY-NLS Variant that Lacks the Proline-Tyrosine Motif. Structure 24:1802-1809
Soniat, Michael; Chook, Yuh Min (2015) Nuclear localization signals for four distinct karyopherin-β nuclear import systems. Biochem J 468:353-62
Fung, Ho Yee Joyce; Fu, Szu-Chin; Brautigam, Chad A et al. (2015) Structural determinants of nuclear export signal orientation in binding to exportin CRM1. Elife 4:
Xu, Darui; Marquis, Kara; Pei, Jimin et al. (2015) LocNES: a computational tool for locating classical NESs in CRM1 cargo proteins. Bioinformatics 31:1357-65
Bernis, Cyril; Swift-Taylor, Beth; Nord, Matthew et al. (2014) Transportin acts to regulate mitotic assembly events by target binding rather than Ran sequestration. Mol Biol Cell 25:992-1009
Fung, Ho Yee Joyce; Chook, Yuh Min (2014) Atomic basis of CRM1-cargo recognition, release and inhibition. Semin Cancer Biol 27:52-61
Soniat, Michael; Sampathkumar, Parthasarathy; Collett, Garen et al. (2013) Crystal structure of human Karyopherin β2 bound to the PY-NLS of Saccharomyces cerevisiae Nab2. J Struct Funct Genomics 14:31-5

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