scholarly journals Archaeal ribosomal proteins possess nuclear localization signal-type motifs: implications for the origin of the cell nucleus

2019 ◽  
Author(s):  
Sergey Melnikov ◽  
Hui-Si Kwok ◽  
Kasidet Manakongtreecheep ◽  
Antonia van den Elzen ◽  
Carson C. Thoreen ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Cell Nucleus ◽  
Ribosomal Proteins ◽  
Eukaryotic Cell ◽  
Nuclear Localization Signals ◽  
Sequence Of Events ◽  
Eukaryotic Proteins ◽  
Signal Type ◽  
Localization Signal ◽  
Short Signal

AbstractEukaryotic cells are divided into the nucleus and the cytosol, and, to enter the nucleus, proteins typically possess short signal sequences, known as nuclear localization signals (NLSs). Although NLSs have long been considered as features unique to eukaryotic proteins, we show here that similar or identical protein segments are present in ribosomal proteins from the Archaea. Specifically, the ribosomal proteins uL3, uL15, uL18, and uS12 possess NLS-type motifs that are conserved across all major branches of the Archaea, including the most ancient groups Microarchaeota and Diapherotrites, pointing to the ancient origin of NLS-type motifs in the Archaea. Furthermore, by using fluorescence microscopy, we show that the archaeal NLS-type motifs can functionally substitute eukaryotic NLSs and direct the transport of ribosomal proteins into the nuclei of human cells. Collectively, these findings illustrate that the origin of NLSs preceded the origin of the cell nucleus, suggesting that the initial function of NLSs was not related to intracellular trafficking. Overall, our study reveals rare evolutionary intermediates among archaeal cells that can help elucidate the sequence of events that led to the origin of the eukaryotic cell.

scholarly journals Archaeal Ribosomal Proteins Possess Nuclear Localization Signal-Type Motifs: Implications for the Origin of the Cell Nucleus

2019 ◽  
Vol 37 (1) ◽  
pp. 124-133 ◽  
Author(s):  
Sergey Melnikov ◽  
Hui-Si Kwok ◽  
Kasidet Manakongtreecheep ◽  
Antonia van den Elzen ◽  
Carson C Thoreen ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Cell Nucleus ◽  
Ribosomal Proteins ◽  
Eukaryotic Cell ◽  
Nuclear Localization Signals ◽  
Sequence Of Events ◽  
Eukaryotic Proteins ◽  
Signal Type ◽  
Localization Signal ◽  
Short Signal

Abstract Eukaryotic cells are divided into the nucleus and the cytosol, and, to enter the nucleus, proteins typically possess short signal sequences, known as nuclear localization signals (NLSs). Although NLSs have long been considered as features unique to eukaryotic proteins, we show here that similar or identical protein segments are present in ribosomal proteins from the Archaea. Specifically, the ribosomal proteins uL3, uL15, uL18, and uS12 possess NLS-type motifs that are conserved across all major branches of the Archaea, including the most ancient groups Microarchaeota and Diapherotrites, pointing to the ancient origin of NLS-type motifs in the Archaea. Furthermore, by using fluorescence microscopy, we show that the archaeal NLS-type motifs can functionally substitute eukaryotic NLSs and direct the transport of ribosomal proteins into the nuclei of human cells. Collectively, these findings illustrate that the origin of NLSs preceded the origin of the cell nucleus, suggesting that the initial function of NLSs was not related to intracellular trafficking, but possibly was to improve recognition of nucleic acids by cellular proteins. Overall, our study reveals rare evolutionary intermediates among archaeal cells that can help elucidate the sequence of events that led to the origin of the eukaryotic cell.

scholarly journals Function of two discrete regions is required for nuclear localization of polymerase basic protein 1 of A/WSN/33 influenza virus (H1 N1).

1990 ◽  
Vol 10 (8) ◽  
pp. 4139-4145 ◽  
Author(s):  
S T Nath ◽  
D P Nayak
Keyword(s):  
Influenza Virus ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Basic Protein ◽  
Localization Property ◽  
Nuclear Localization Signals ◽  
Chicken Muscle ◽  
Localization Signal ◽  
Cloned Cdna ◽  
Muscle Pyruvate Kinase

Polymerase basic protein 1 (PB1) of influenza virus (A/WSN/33), when expressed from cloned cDNA in the absence of other viral proteins, accumulates in the nucleus. We have examined the location and nature of the nuclear localization signal of PB1 by using deletion mutants and chimeric constructions with chicken muscle pyruvate kinase, a cytoplasmic protein. Our studies showed some novel features of the nuclear localization signal of PB1. The signal was present internally within residues 180 to 252 of PB1. Moreover, unlike most nuclear localization signals, it was not a single stretch of contiguous amino acids. Instead, it possessed two discontinuous regions separated by an intervening sequence which could be deleted without affecting its nuclear localization property. On the other hand, deletion of either of the two signal regions rendered the protein cytoplasmic, indicating that the function of both regions is required for nuclear localization and that one region alone is not sufficient. Both of these signal regions contained short stretches of basic residues. Possible ways by which this novel bipartite signal can function in nuclear localization are discussed.

One pot synthesis of highly luminescent polyethylene glycol anchored carbon dots functionalized with a nuclear localization signal peptide for cell nucleus imaging

Nanoscale ◽  
2015 ◽  
Vol 7 (14) ◽  
pp. 6104-6113 ◽  
Author(s):  
Lei Yang ◽  
Weihua Jiang ◽  
Lipeng Qiu ◽  
Xuewei Jiang ◽  
Daiying Zuo ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Nuclear Localization ◽  
Signal Peptide ◽  
Cell Nucleus ◽  
Nuclear Localization Signal ◽  
Carbon Dots ◽  
One Pot ◽  
One Pot Synthesis ◽  
Localization Signal

Mutations of the SBDS Gene Result in Nuclear Mislocalization.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2034-2034
Author(s):  
Masafumi Yamaguchi ◽  
Kingo Fujimura ◽  
Hanae Toga-Yamaguchi ◽  
Valentina Svetic ◽  
Naoki Okamura ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Bone Marrow Failure ◽  
Pancreatic Insufficiency ◽  
Exocrine Pancreatic Insufficiency ◽  
Expression Level ◽  
Wild Type ◽  
Nuclear Localization Signals ◽  
Localization Signal ◽  
Domain I

Abstract Shwachman-Diamond syndrome (SDS) is an autosomal-recessive disorder characterized by exocrine pancreatic insufficiency and bone marrow failure. The SDS disease locus was mapped to chromosome 7q11. We have previously reported that Shwachman-Bodian- Diamond syndrome (SBDS) gene is not required for neutrophil maturation. However, SBDS knockdown cells were sensitive to apoptotic stimuli, indicating that SBDS acts to maintain survival of granulocyte precursor cells. (Exp Hematol35; 579, 2007). A wide variety of mutations in SBDS gene has been identified, and almost of all patients show truncated immature proteins, p.K62X (c.183_184TA>CT) or p.C84fsX3 (c.258+2T>C). However, it is not yet clear how these truncated proteins affect cellular processes that result in the SDS phenotype. The SBDS protein is localized to the nucleoli but does not have the canonical nuclear localization signal. In order to clarify the molecular basis of pathogenicity of mutated SBDS proteins, we explored the subcellular distribution of normal and mutant SBDS proteins in Hela and 32Dcl3 cells. Using various N-terminal and C-terminal deletion constructs, we found N-terminal region, domain I (1-87 amino acid residue) in particular, was necessary to localize to the nucleus. The disease related mutations (C31W, K33E, N34I, L71P) and the mutations which are conserved among the species in the domain I (E44K, K62E, D70N, E82K) were generated. C31W and N34I mutants failed to localize SBDS to the nuclei. The SV40 derived nuclear localization signal was fused to these mutated SBDS protein, and these proteins were clearly localized to the nuclei. In addition to the mislocalization, the protein expression level of these mutants showed a dramatic decrease compared to the wild type. We also established SBDS wild type and domain I overexpressed 32Dcl3 cell. SBDS wild type overexpressed cells could differentiate to normal neutrophils in the presence of mG-CSF, however domain I overexpressed cells did not differentiate. Almost of all cells showed apoptosis in this domain I overexpressed cells in the presence of mG-CSF, and this was very similar like SBDS RNAi knockdown cells. The localization of endogenous SBDS protein was also analyzed in this domain I overexpressed cells. The domain I was concentrated to nuclei, however endogenous SBDS protein was diffused to cytosol. Conclusions: The present findings enable us to document the nuclear localization signals in SBDS domain I, and that the shuttling protein would promote SBDS to nuclei. These results also showed that mislocalization and/or low expression level of mutated SBDS protein would cause SDS.

scholarly journals Phosphorylation adjacent to the nuclear localization signal of human dUTPase abolishes nuclear import: structural and mechanistic insights

2013 ◽  
Vol 69 (12) ◽  
pp. 2495-2505 ◽  
Author(s):  
Gergely Róna ◽  
Mary Marfori ◽  
Máté Borsos ◽  
Ildikó Scheer ◽  
Enikő Takács ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Nucleocytoplasmic Transport ◽  
Wild Type ◽  
Post Translational Modification ◽  
Nuclear Localization Signals ◽  
M Phase ◽  
Importin Α ◽  
Localization Signal

Phosphorylation adjacent to nuclear localization signals (NLSs) is involved in the regulation of nucleocytoplasmic transport. The nuclear isoform of human dUTPase, an enzyme that is essential for genomic integrity, has been shown to be phosphorylated on a serine residue (Ser11) in the vicinity of its nuclear localization signal; however, the effect of this phosphorylation is not yet known. To investigate this issue, an integrated set of structural, molecular and cell biological methods were employed. It is shown that NLS-adjacent phosphorylation of dUTPase occurs during the M phase of the cell cycle. Comparison of the cellular distribution of wild-type dUTPase with those of hyperphosphorylation- and hypophosphorylation-mimicking mutants suggests that phosphorylation at Ser11 leads to the exclusion of dUTPase from the nucleus. Isothermal titration microcalorimetry and additional independent biophysical techniques show that the interaction between dUTPase and importin-α, the karyopherin molecule responsible for `classical' NLS binding, is weakened significantly in the case of the S11E hyperphosphorylation-mimicking mutant. The structures of the importin-α–wild-type and the importin-α–hyperphosphorylation-mimicking dUTPase NLS complexes provide structural insights into the molecular details of this regulation. The data indicate that the post-translational modification of dUTPase during the cell cycle may modulate the nuclear availability of this enzyme.

scholarly journals Human mRNA Cap Methyltransferase: Alternative Nuclear Localization Signal Motifs Ensure Nuclear Localization Required for Viability

2005 ◽  
Vol 25 (7) ◽  
pp. 2644-2649 ◽  
Author(s):  
Beth Shafer ◽  
Chun Chu ◽  
Aaron J. Shatkin
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Nuclear Localization Signals ◽  
Truncation Mutant ◽  
Capping Enzyme ◽  
Importin Α ◽  
Localization Signal ◽  
Targeting Signals ◽  
Sirna Knockdown ◽  
Interfering Rna

ABSTRACT A characteristic feature of gene expression in eukaryotes is the addition of a 5′-terminal 7-methylguanine cap (m7GpppN) to nascent pre-mRNAs in the nucleus catalyzed by capping enzyme and cap methyltransferase. Small interfering RNA (siRNA) knockdown of cap methyltransferase in HeLa cells resulted in apoptosis as measured by terminal deoxynucleotidyltransferase-mediated dUTP-tetramethylrhodamine nick end labeling assay, demonstrating the importance of mRNA 5′-end methylation for mammalian cell viability. Nuclear localization of cap methyltransferase is mediated by interaction with importin-α, which facilitates its transport and selective binding to transcripts containing 5′-terminal GpppN. The methyltransferase 96-144 region has been shown to be necessary for importin binding, and N-terminal fusion of this sequence to nonnuclear proteins proved sufficient for nuclear localization. The targeting sequence was narrowed to amino acids 120 to 129, including a required 126KRK. Although full-length methyltransferase (positions 1 to 476) contains the predicted nuclear localization signals 57RKRK, 80KKRK, 103KKRKR, and 194KKKR, mutagenesis studies confirmed functional motifs only at positions 80, 103, and the previously unrecognized 126KRK. All three motifs can act as alternative nu clear targeting signals. Expression of N-truncated cap methyltransferase (120 to 476) restored viability of methyltransferase siRNA knocked-down cells. However, an enzymatically active 144-476 truncation mutant missing the three nuclear localization signals was mostly cytoplasmic and ineffective in preventing siRNA-induced loss of viability.

scholarly journals Localization and Functional Roles of Components of the Translation Apparatus in the Eukaryotic Cell Nucleus

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3239
Author(s):  
Zaur M. Kachaev ◽  
Sergey D. Ivashchenko ◽  
Eugene N. Kozlov ◽  
Lyubov A. Lebedeva ◽  
Yulii V. Shidlovskii
Keyword(s):  
Gene Expression ◽  
Cell Nucleus ◽  
Ribosomal Proteins ◽  
Nuclear Translocation ◽  
Eukaryotic Cell ◽  
Current Data ◽  
Cell Nuclei ◽  
Functional Roles ◽  
Translation Apparatus ◽  
Cell Stress Response

Components of the translation apparatus, including ribosomal proteins, have been found in cell nuclei in various organisms. Components of the translation apparatus are involved in various nuclear processes, particularly those associated with genome integrity control and the nuclear stages of gene expression, such as transcription, mRNA processing, and mRNA export. Components of the translation apparatus control intranuclear trafficking; the nuclear import and export of RNA and proteins; and regulate the activity, stability, and functional recruitment of nuclear proteins. The nuclear translocation of these components is often involved in the cell response to stimulation and stress, in addition to playing critical roles in oncogenesis and viral infection. Many components of the translation apparatus are moonlighting proteins, involved in integral cell stress response and coupling of gene expression subprocesses. Thus, this phenomenon represents a significant interest for both basic and applied molecular biology. Here, we provide an overview of the current data regarding the molecular functions of translation factors and ribosomal proteins in the cell nucleus.

scholarly journals Effect of Different Nuclear Localization Signals on the Subcellular Localization and Anti-HIV-1 Function of the MxB Protein

2021 ◽  
Vol 12 ◽  
Author(s):  
Keli Chai ◽  
Zhen Wang ◽  
Qinghua Pan ◽  
Juan Tan ◽  
Wentao Qiao ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Nuclear Import ◽  
Molecular Mechanisms ◽  
Virus Production ◽  
Viral Capsid ◽  
Terminal Sequence ◽  
Nuclear Localization Signals ◽  
Localization Signal ◽  
Anti Hiv ◽  
Hiv 1

Interferon exerts its antiviral activity by stimulating the expression of antiviral proteins. These interferon stimulate genes (ISGs) often target a group of viruses with unique molecular mechanisms. One such ISG is myxovirus resistance B (MxB) that has been reported to inhibit human immunodeficiency virus type 1 (HIV-1) by targeting viral capsid and impairing nuclear import of viral DNA. The antiviral specificity of MxB is determined by its N-terminal 25 amino acids sequence which has the nuclear localization activity, therefore functions as a nuclear localization signal (NLS). In this study, we report that the bipartite NLS, but not the classic NLS, the PY-NLS, nor the arginine-rich NLS, when used to replace the N-terminal sequence of MxB, drastically suppress HIV-1 gene expression and virus production, thus creates a new anti-HIV-1 mechanism. MxB preserves its anti-HIV-1 activity when its N-terminal sequence is replaced by the arginine-rich NLS. Interestingly, the arginine-rich NLS allows MxB to inhibit HIV-1 CA mutants that are otherwise resistant to wild type MxB, which suggests sequence specific targeting of viral capsid. Together, these data implicate that it is not the nuclear import function itself, but rather the sequence and the mechanism of action of the NLS which define the antiviral property of MxB.

scholarly journals Origin of the nuclear proteome on the basis of pre-existing nuclear localization signals in prokaryotic proteins

2020 ◽  
Author(s):  
Olga M. Lisitsyna ◽  
Margarita A. Kurnaeva ◽  
Eugene A. Arifulin ◽  
Maria Y. Shubina ◽  
Yana R. Musinova ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Protein Import ◽  
Eukaryotic Cell ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Nuclear Localization Signals ◽  
Binding Domains ◽  
Nuclear Proteome ◽  
Pore Complexes ◽  
Open Question

AbstractThe origin of the selective nuclear protein import machinery, which consists of nuclear pore complexes and adaptor molecules interacting with the nuclear localization signals (NLSs) of cargo molecules, was one of the most important events in the evolution of the eukaryotic cell. How the proteins were selected for import into the forming nuclei remains an open question. Here, we demonstrate that functional NLSs may be integrated inside nucleotide-binding domains of both eukaryotic and prokaryotic proteins and may co-evolve with these domains. We propose that the pre-existence of NLSs inside prokaryotic proteins dictated, at least partially, the nuclear proteome composition.

scholarly journals Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus

1999 ◽  
Vol 96 (1) ◽  
pp. 91-96 ◽  
Author(s):  
M. A. Zanta ◽  
P. Belguise-Valladier ◽  
J.-P. Behr
Keyword(s):  
Gene Delivery ◽  
Nuclear Localization ◽  
Signal Peptide ◽  
Cell Nucleus ◽  
Nuclear Localization Signal ◽  
Localization Signal
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