Phosphorylation in the accessory domain of yeast histone chaperone protein 1 exposes the nuclear export signal sequence

10.22541/au.161550022.24032527/v1 ◽

2021 ◽

Author(s):

Sho Ashida ◽

Rikuri Morita ◽

Yasuteru Shigeta ◽

Ryuhei Harada

Keyword(s):

Cell Nucleus ◽

Nuclear Export ◽

Signal Sequence ◽

Nuclear Export Signal ◽

Md Simulations ◽

Histone Chaperone ◽

Chaperone Protein ◽

Accessory Domain ◽

Export Signal

Histone is a scaffold protein that constitutes nucleosomes with DNA in the cell nucleus. When forming histone, hetero octamer is assisted by histone chaperone proteins. As a histone chaperone protein, the crystal structure of yeast nucleosome assembly protein (yNap1) has been determined. For yNap1, a nuclear export signal/sequence (NES) has been identified as a part of the long -helix. Experimental evidence via mutagenesis on budding yeast suggests the NES is necessary for transport out from the cell nucleus. However, the NES is masked by a region defined as an accessory domain (AD). In addition, the role of the AD in nuclear transport has not been elucidated yet. To address the role of the AD, we focused on phosphorylation in the AD because proteome experiments have identified multiple phosphorylation sites of yNap1. To computationally treat phosphorylation, we performed all-atom molecular dynamics (MD) simulations for a set of non-phosphorylated and phosphorylated yNap1 (Nap1-nonP and Nap1-P). As an analysis, we addressed how the NES is exposed to the protein surface by measuring its solvent-access surface area (SASA). As a result, there was a difference in the SASA distributions between both systems. Quantitatively, the median of the SASA distribution of Nap1-P was greater than that of Nap1-nonP, meaning that phosphorylation in the AD exposed to the NES, resulting in increasing its accessibility. In conclusion, yNap1 might modulate the accessibility of the NES by dislocating the AD through phosphorylation.

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Nuclear Import of Hepatic Glucokinase Depends upon Glucokinase Regulatory Protein, whereas Export Is Due to a Nuclear Export Signal Sequence in Glucokinase

Journal of Biological Chemistry ◽

10.1074/jbc.274.52.37125 ◽

1999 ◽

Vol 274 (52) ◽

pp. 37125-37130 ◽

Cited By ~ 97

Author(s):

Chiyo Shiota ◽

Jack Coffey ◽

Joseph Grimsby ◽

Joseph F. Grippo ◽

Mark A. Magnuson

Keyword(s):

Nuclear Import ◽

Nuclear Export ◽

Signal Sequence ◽

Regulatory Protein ◽

Nuclear Export Signal ◽

Glucokinase Regulatory Protein ◽

Export Signal

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Identification of a nuclear export signal sequence for bovine papillomavirus E1 protein

Virology ◽

10.1016/j.virol.2007.12.017 ◽

2008 ◽

Vol 373 (1) ◽

pp. 149-162 ◽

Cited By ~ 13

Author(s):

Germán Rosas-Acosta ◽

Van G. Wilson

Keyword(s):

Nuclear Export ◽

Signal Sequence ◽

Nuclear Export Signal ◽

Bovine Papillomavirus ◽

E1 Protein ◽

Export Signal

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Generation of Influenza A Virus NS2 (NEP) Mutants with an Altered Nuclear Export Signal Sequence

Journal of Virology ◽

10.1128/jvi.78.18.10149-10155.2004 ◽

2004 ◽

Vol 78 (18) ◽

pp. 10149-10155 ◽

Cited By ~ 33

Author(s):

Kiyoko Iwatsuki-Horimoto ◽

Taisuke Horimoto ◽

Yutaka Fujii ◽

Yoshihiro Kawaoka

Keyword(s):

Influenza A Virus ◽

Nuclear Export ◽

Influenza A ◽

Signal Sequence ◽

Nuclear Export Signal ◽

Wild Type Virus ◽

Wild Type ◽

Amino Terminal ◽

Ribonucleoprotein Complexes ◽

Export Signal

ABSTRACT The NS2 (NEP) protein of influenza A virus contains a highly conserved nuclear export signal (NES) motif in its amino-terminal region (12ILMRMSKMQL21, A/WSN/33), which is thought to be required for nuclear export of viral ribonucleoprotein complexes (vRNPs) mediated by a cellular export factor, CRM1. However, simultaneous replacement of three hydrophobic residues in the NES with alanine does not affect NS2 (NEP) binding to CRM1, although the virus with these mutations is not viable. To determine the extent of sequence conservation required by the NS2 (NEP) NES for its export function during viral replication, we randomly introduced mutations by degenerative mutagenesis into the region of NS cDNA encoding the NS2 (NEP) NES and then attempted to generate mutant viruses containing these alterations by reverse genetics. Sequence analysis of the recovered viruses showed that although some of the mutants possessed amino acids other than those conserved in the NES, hydrophobicity within this motif was maintained. Nuclear export of vRNPs representing all of the mutant viruses was completely inhibited in the presence of a CRM1 inhibitor, leptomycin B, as was the transport of wild-type virus, indicating that the CRM1-mediated pathway is responsible for the nuclear export of both wild-type and mutant vRNPs. The vRNPs of some of the mutant viruses were exported in a delayed manner, resulting in limited viral growth in cell culture and in mice. These results suggest that the NES motif may be an attractive target for the introduction of attenuating mutations in the production of live vaccine viruses.

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A factor required for nonsense-mediated mRNA decay in yeast is exported from the nucleus to the cytoplasm by a nuclear export signal sequence

Journal of Cell Science ◽

10.1242/jcs.111.21.3129 ◽

1998 ◽

Vol 111 (21) ◽

pp. 3129-3143 ◽

Cited By ~ 4

Author(s):

R.L. Shirley ◽

M.J. Lelivelt ◽

L.R. Schenkman ◽

J.N. Dahlseid ◽

M.R. Culbertson

Keyword(s):

Protein Function ◽

Nuclear Export ◽

Mrna Decay ◽

Signal Sequence ◽

Nuclear Export Signal ◽

Reporter Protein ◽

Nuclear Localization Signals ◽

Sequence Elements ◽

Nonsense Mediated Mrna Decay ◽

Export Signal

In Saccharomyces cerevisiae, Upf3p is required for nonsense-mediated mRNA decay (NMD). Although localized primarily in the cytoplasm, Upf3p contains three sequence elements that resemble nuclear localization signals (NLSs) and two sequence elements that resemble nuclear export signals (NESs). We found that a cytoplasmic reporter protein localized to the nucleus when fused to any one of the three NLS-like sequences of Upf3p. A nuclear reporter protein localized to the cytoplasm when fused to one of the NES-like sequences (NES-A). We present evidence that NES-A functions to signal the export of Upf3p from the nucleus. Combined alanine substitutions in the NES-A element caused a re-distribution of Upf3p to a subnuclear location identified as the nucleolus and conferred an Nmd- phenotype. Single mutations in NES-A failed to affect the distribution of Upf3p and were Nmd+. When an NES element from HIV-1 Rev was inserted near the C terminus of a mutant Upf3p containing multiple mutations in NES-A, the cytoplasmic distribution typical of wild-type Upf3p was restored but the cells remained phenotypically Nmd-. These results suggest that NES-A is a functional nuclear export signal. Combined mutations in NES-A may cause multiple defects in protein function leading to an Nmd- phenotype even when export is restored.

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Phospholipase C-δ1 Contains a Functional Nuclear Export Signal Sequence

Journal of Biological Chemistry ◽

10.1074/jbc.274.40.28537 ◽

1999 ◽

Vol 274 (40) ◽

pp. 28537-28541 ◽

Cited By ~ 56

Author(s):

Masaki Yamaga ◽

Makoto Fujii ◽

Hideaki Kamata ◽

Hajime Hirata ◽

Hitoshi Yagisawa

Keyword(s):

Phospholipase C ◽

Nuclear Export ◽

Signal Sequence ◽

Nuclear Export Signal ◽

Export Signal

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PDZ domain from Dishevelled -- a specificity study.

Acta Biochimica Polonica ◽

10.18388/abp.2011_2272 ◽

2011 ◽

Vol 58 (2) ◽

Cited By ~ 3

Author(s):

Katarzyna Śmietana ◽

Agnieszka Mateja ◽

Artur Krężel ◽

Jacek Otlewski

Keyword(s):

Wnt Signaling ◽

Nuclear Export ◽

Signal Sequence ◽

Consensus Sequence ◽

Pdz Domain ◽

Nuclear Export Signal ◽

Signaling Cascades ◽

Binding Motif ◽

Wnt Proteins ◽

Export Signal

Intracellular signaling cascades induced by Wnt proteins play a key role in developmental processes and are implicated in cancerogenesis. It is still unclear how the cell determines which of the three possible Wnt response mechanisms should be activated, but the decision process is most likely dependent on Dishevelled proteins. Dishevelled family members interact with many diverse targets, however, molecular mechanisms underlying these binding events have not been comprehensively described so far. Here, we investigated the specificity of the PDZ domain from human Dishevelled-2 using C-terminal phage display, which led us to identification of a leucine-rich binding motif strongly resembling the consensus sequence of a nuclear export signal. PDZ interactions with several peptide and protein motifs (including the nuclear export signal sequence from Dishevelled-2 protein) were investigated in detail using fluorescence spectroscopy, mutational analysis and immunoenzymatic assays. The experiments showed that the PDZ domain can bind the nuclear export signal sequence of the Dishevelled-2 protein. Since the intracellular localization of Dishevelled is governed by nuclear localization and nuclear export signal sequences, it is possible that the intramolecular interaction between PDZ domain and the export signal could modulate the balance between nuclear and cytoplasmic pool of the Dishevelled protein. Such a regulatory mechanism would be of utmost importance for the differential activation of Wnt signaling cascades, leading to selective promotion of the nucleus-dependent Wnt β-catenin pathway at the expense of non-canonical Wnt signaling.

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