PDBsum entry 1y2a

Protein transport

PDB id

1y2a

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Contents

			Protein chain

					423 a.a. *

			Ligands

					GLY-LYS-ILE-SER- LYS-HIS-TRP-THR- GLY-ILE

			Waters ×134

* Residue conservation analysis

PDB id:

1y2a

Links

Name:

Protein transport

Title:

Structure of mammalian importin bound to the non-classical plscr1-nls

Structure:

Importin alpha-2 subunit. Chain: c. Fragment: residues 70-497. Synonym: karyopherin alpha-2 subunit. Srp1-alpha. Rag cohort protein 1. Pendulin. Pore targeting complex 58 kda subunit. Ptac58. Importin alpha p1. Engineered: yes. Decamer fragment of phospholipid scramblase 1. Chain: p.

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Gene: kpna2, rch1. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Synthetic: yes. Other_details: this sequence occurs naturally in homo sapiens, gene plscr1

Biol. unit:

Dimer (from PQS)

Resolution:

2.20Å

R-factor:

0.229

R-free:

0.258

Authors:

M.-H.Chen,I.Ben-Efraim,G.Mitrousis,N.Walker-Kopp,P.J.Sims,G.Cingolani

Key ref:

M.H.Chen et al. (2005). Phospholipid scramblase 1 contains a nonclassical nuclear localization signal with unique binding site in importin alpha. J Biol Chem, 280, 10599-10606. PubMed id: 15611084 DOI: 10.1074/jbc.M413194200

Date:

22-Nov-04

Release date:

01-Feb-05

PROCHECK

	Headers

	References

Protein chain

P52293 (IMA1_MOUSE) - Importin subunit alpha-1 from Mus musculus

Seq: Struc:

Seq: Struc:					529 a.a. 423 a.a.

Key:

PfamA domain

Secondary structure

CATH domain

DOI no: 10.1074/jbc.M413194200	J Biol Chem 280:10599-10606 (2005)
PubMed id: 15611084

Phospholipid scramblase 1 contains a nonclassical nuclear localization signal with unique binding site in importin alpha.
M.H.Chen, I.Ben-Efraim, G.Mitrousis, N.Walker-Kopp, P.J.Sims, G.Cingolani.

ABSTRACT

Nuclear import of proteins containing a classical nuclear localization signal (NLS) is an energy-dependent process that requires the heterodimer importin alpha/beta. Three to six basic contiguous arginine/lysine residues characterize a classical NLS and are thought to form a basic patch on the surface of the import cargo. In this study, we have characterized the NLS of phospholipid scramblase 1 (PLSCR1), a lipid-binding protein that enters the nucleus via the nonclassical NLS (257)GKISKHWTGI(266). This import sequence lacks a contiguous stretch of positively charged residues, and it is enriched in hydrophobic residues. We have determined the 2.2 A crystal structure of a complex between the PLSCR1 NLS and the armadillo repeat core of vertebrate importin alpha. Our crystallographic analysis reveals that PLSCR1 NLS binds to armadillo repeats 1-4 of importin alpha, but its interaction partially overlaps the classical NLS binding site. Two PLSCR1 lysines occupy the canonical positions indicated as P2 and P5. Moreover, we present in vivo evidence that the critical lysine at position P2, which is essential in other known NLS sequences, is dispensable in PLSCR1 NLS. Taken together, these data provide insight into a novel nuclear localization signal that presents a distinct motif for binding to importin alpha.

Selected figure(s)



	Figure 3. FIG. 3. Analysis of the IBB-importin -PLSCR1 NLS binding interface. A, electrostatic surface potential of importin . Acidic and basic surfaces of the protein are in red and blue, respectively. The PLSCR1 NLS, in blue sticks, is docked to the importin surface. B, recognition of PLSCR1 Lys261. The conserved lysine at position P5 is sandwiched by two importin Trp184/ Trp142 (shown in yellow) and by the PLSCR1-Trp263 (in orange).		Figure 5. FIG. 5. Nuclear import of PLSCR1 is blocked by triple mutation of the residues K258A, K261A/H263A, but not by single point mutations. Confocal fluorescence images of murine SVT2 fibroblasts transiently transfected with (top to bottom): pcDNA3 containing cDNA for wild-type (WT) human PLSCR1 or the mutants PLSCR1 (184AAAPAA^189) or PLSCR1 (184AAAPAA^189, K258A), or (184-AAAPAA^189, K261A) or PLSCR1(184AAAPAA^189, W263A) or (184AAAPAA^189, 258-GAISAAWTGI266). First column PL-SCR1 label, second column nuclear label, third column merge. Expressed human PLSCR1 antigen was detected with monoclonal antibody 4D2 and fluorescein isothiocyanate-conjugated goat anti-mouse IgG. Following immunostaining and DNA labeling with propidium iodide (PI), cells were visualized by confocal fluorescence microscopy. Data are from a single experiment, representative of four so performed.

Figures were selected by an automated process.

Literature references that cite this PDB file's key reference

PubMed id

Reference

20028483

A.Lange, L.M.McLane, R.E.Mills, S.E.Devine, and A.H.Corbett (2010).
Expanding the definition of the classical bipartite nuclear localization signal.

Traffic, 11, 311-323.

20412059

G.Merényi, E.Kónya, and B.G.Vértessy (2010).
Drosophila proteins involved in metabolism of uracil-DNA possess different types of nuclear localization signals.

FEBS J, 277, 2142-2156.

20435918

N.Gudleski, J.M.Flanagan, E.P.Ryan, M.C.Bewley, and L.J.Parent (2010).
Directionality of nucleocytoplasmic transport of the retroviral gag protein depends on sequential binding of karyopherins and viral RNA.

Proc Natl Acad Sci U S A, 107, 9358-9363.

19333378

B.Py, S.Basmaciogullari, J.Bouchet, M.Zarka, I.C.Moura, M.Benhamou, R.C.Monteiro, H.Hocini, R.Madrid, and S.Benichou (2009).
The phospholipid scramblases 1 and 4 are cellular receptors for the secretory leukocyte protease inhibitor and interact with CD4 at the plasma membrane.

PLoS ONE, 4, e5006.

19514019

L.M.McLane, and A.H.Corbett (2009).
Nuclear localization signals and human disease.

IUBMB Life, 61, 697-706.

19244342

Z.Shen, N.Paquin, A.Forget, and P.Chartrand (2009).
Nuclear shuttling of She2p couples ASH1 mRNA localization to its translational repression by recruiting Loc1p and Puf6p.

Mol Biol Cell, 20, 2265-2275.

17682055

M.M.Pradeepa, S.Manjunatha, V.Sathish, S.Agrawal, and M.R.Rao (2008).
Involvement of importin-4 in the transport of transition protein 2 into the spermatid nucleus.

Mol Cell Biol, 28, 4331-4341.

17359931

G.I.Belogrudov (2007).
A 24-residue presequence localizes human factor B to mitochondria.

Arch Biochem Biophys, 461, 95.

17567603

J.P.Wyles, Z.Wu, S.E.Mirski, and S.P.Cole (2007).
Nuclear interactions of topoisomerase II alpha and beta with phospholipid scramblase 1.

Nucleic Acids Res, 35, 4076-4085.

17118134

A.Kel, N.Voss, R.Jauregui, O.Kel-Margoulis, and E.Wingender (2006).
Beyond microarrays: Finding key transcription factors controlling signal transduction pathways.

BMC Bioinformatics, 7, S13.

16421734

A.S.Madrid, and K.Weis (2006).
Nuclear transport is becoming crystal clear.

Chromosoma, 115, 98.

17254297

H.H.Lin, L.Y.Han, H.L.Zhang, C.J.Zheng, B.Xie, Z.W.Cao, and Y.Z.Chen (2006).
Prediction of the functional class of metal-binding proteins from sequence derived physicochemical properties by support vector machine approach.

BMC Bioinformatics, 7, S13.

16702944

Y.Huang, Q.Zhao, C.X.Zhou, Z.M.Gu, D.Li, H.Z.Xu, T.Wiedmer, P.J.Sims, K.W.Zhao, and G.Q.Chen (2006).
Antileukemic roles of human phospholipid scramblase 1 gene, evidence from inducible PLSCR1-expressing leukemic cells.

Oncogene, 25, 6618-6627.

16611984

Y.Li, K.Rogulski, Q.Zhou, P.J.Sims, and E.V.Prochownik (2006).
The negative c-Myc target onzin affects proliferation and apoptosis via its obligate interaction with phospholipid scramblase 1.

Mol Cell Biol, 26, 3401-3413.

The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.