Plant β-tubulin phosphorylation on Ser172 as canonical suppressing factor of microtubule growth

Aim. The estimation of potential role of plant β-tubulin Ser172 phosphorylation for correct function of microtubules and cell division due to selection of protein kinases, most probable associated with phosphorylation of Ser172 in Arabidopsis thaliana (L.) Heynh. Methods. Literature and database search. Comparison of protein sequences and structures: multiple sequence alignment, phylogenetic profiling, protein structure modeling, etc. Results. Comparison of Ser172 site region from all known β-tubulins from Homo sapiens, Sus scrofa, Saccharomyces cerevisiae, Drosophila melanogaster and A. thaliana confirms its significant similarity. Joint clusterization of all Ser172 site regions (in S±10 a.a. format) reveals that plant site is most similar to Ser172±10 fragment of β-tubulin from S. cerevisiae. At the same time, sequences and catalytic domain structures of cyclin-dependent kinases 1 and YAK1-related kinases (MNB/DYRK1a/YAK1) associated with Ser172 phosphorylation, found maximal similarity in A. thaliana and S. cerevisiae. Сonclusions. The results confirm similarity of amino acid environment of Ser172 in β-tubulin isotypes in human, pig, fruit fly, yeast and arabidopsis. This suggests similar effect of β-tubulin phosphorylation at Ser172 for inhibition of microtubule assembly onto their protofilaments and its association with CDK1 and YAK1-related protein kinases. Similarity of Ser172 sites and associated protein kinases, allows us to expect similar effect of this modification on structure of microtubules in A. thaliana and S. cerevisiae. Keywords: β-tubulin, Ser172, phosphorylation, CDK1, DYRK1, MNB, YAK1.

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Molecular and Comparative Genetics of Mental Retardation

Genetics ◽

10.1093/genetics/166.2.835 ◽

2004 ◽

Vol 166 (2) ◽

pp. 835-881 ◽

Cited By ~ 2

Author(s):

Jennifer K Inlow ◽

Linda L Restifo

Keyword(s):

Mental Retardation ◽

Homo Sapiens ◽

System Development ◽

Laboratory Data ◽

Fruit Fly ◽

Mendelian Inheritance ◽

Nervous System Development ◽

Skewed Distribution ◽

Therapeutic Drug ◽

Cellular Mechanisms

Abstract Affecting 1-3% of the population, mental retardation (MR) poses significant challenges for clinicians and scientists. Understanding the biology of MR is complicated by the extraordinary heterogeneity of genetic MR disorders. Detailed analyses of >1000 Online Mendelian Inheritance in Man (OMIM) database entries and literature searches through September 2003 revealed 282 molecularly identified MR genes. We estimate that hundreds more MR genes remain to be identified. A novel test, in which we distributed unmapped MR disorders proportionately across the autosomes, failed to eliminate the well-known X-chromosome overrepresentation of MR genes and candidate genes. This evidence argues against ascertainment bias as the main cause of the skewed distribution. On the basis of a synthesis of clinical and laboratory data, we developed a biological functions classification scheme for MR genes. Metabolic pathways, signaling pathways, and transcription are the most common functions, but numerous other aspects of neuronal and glial biology are controlled by MR genes as well. Using protein sequence and domain-organization comparisons, we found a striking conservation of MR genes and genetic pathways across the ∼700 million years that separate Homo sapiens and Drosophila melanogaster. Eighty-seven percent have one or more fruit fly homologs and 76% have at least one candidate functional ortholog. We propose that D. melanogaster can be used in a systematic manner to study MR and possibly to develop bioassays for therapeutic drug discovery. We selected 42 Drosophila orthologs as most likely to reveal molecular and cellular mechanisms of nervous system development or plasticity relevant to MR.

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The conserved lysine of the catalytic domain of protein kinases is actively involved in the phosphotransfer reaction and not required for anchoring ATP.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.90.2.442 ◽

1993 ◽

Vol 90 (2) ◽

pp. 442-446 ◽

Cited By ~ 127

Author(s):

A. C. Carrera ◽

K. Alexandrov ◽

T. M. Roberts

Keyword(s):

Protein Kinases ◽

Catalytic Domain

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Open Reading Frame sso2387 from the Archaeon Sulfolobus solfataricus Encodes a Polypeptide with Protein-Serine Kinase Activity

Journal of Bacteriology ◽

10.1128/jb.185.11.3436-3445.2003 ◽

2003 ◽

Vol 185 (11) ◽

pp. 3436-3445 ◽

Cited By ~ 25

Author(s):

Brian H. Lower ◽

Peter J. Kennelly

Keyword(s):

Protein Kinases ◽

Catalytic Domain ◽

Sulfolobus Solfataricus ◽

Open Reading Frame ◽

Serine Kinase ◽

Eukaryotic Protein Kinase ◽

Reading Frame ◽

Eukaryotic Protein ◽

High Concentrations ◽

Eukaryotic Protein Kinases

ABSTRACT The predicted polypeptide product of open reading frame sso2387 from the archaeon Sulfolobus solfataricus, SsoPK2, displayed several of the sequence features conserved among the members of the “eukaryotic” protein kinase superfamily. sso2387 was cloned, and its polypeptide product was expressed in Escherichia coli. The recombinant protein, rSsoPK2, was recovered in insoluble aggregates that could be dispersed by using high concentrations (5 M) of urea. The solubilized polypeptide displayed the ability to phosphorylate itself as well as several exogenous proteins, including mixed histones, casein, bovine serum albumin, and reduced carboxyamidomethylated and maleylated lysozyme, on serine residues. The source of this activity resided in that portion of the protein displaying homology to the catalytic domain of eukaryotic protein kinases. By use of mass spectrometry, the sites of autophosphorylation were found to be located in two areas, one immediately N terminal to the region corresponding to subdomain I of eukaryotic protein kinases, and the second N terminal to the presumed activation loop located between subdomains VII and VIII. Autophosphorylation of rSsoPK2 could be uncoupled from the phosphorylation of exogenous proteins by manipulation of the temperature or mutagenic alteration of the enzyme. Autophosphorylation was detected only at temperatures ≥60°C, whereas phosphorylation of exogenous proteins was detectable at 37°C. Similarly, replacement of one of the potential sites of autophosphorylation, Ser548, with alanine blocked autophosphorylation but not phosphorylation of an exogenous protein, casein.

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Identification and characterization of a novel sucrose-non-fermenting protein kinase/AMP-activated protein kinase-related protein kinase, SNARK

Biochemical Journal ◽

10.1042/bj3550297 ◽

2001 ◽

Vol 355 (2) ◽

pp. 297-305 ◽

Cited By ~ 25

Author(s):

Diana L. LEFEBVRE ◽

Yahong BAI ◽

Nazanin SHAHMOLKY ◽

Monika SHARMA ◽

Raymond POON ◽

...

Keyword(s):

Protein Kinase ◽

Cellular Response ◽

Catalytic Domain ◽

Metabolic Stress ◽

Glucose Deprivation ◽

Protein Band ◽

Northern Blotting ◽

Significant Similarity ◽

Amp Activated Protein Kinase

Subtraction hybridization after the exposure of keratinocytes to ultraviolet radiation identified a differentially expressed cDNA that encodes a protein of 630 amino acid residues possessing significant similarity to the catalytic domain of the sucrose-non-fermenting protein kinase (SNF1)/AMP-activated protein kinase (AMPK) family of serine/threonine protein kinases. Northern blotting and reverse-transcriptase-mediated PCR demonstrated that mRNA transcripts for the SNF1/AMPK-related kinase (SNARK) were widely expressed in rodent tissues. The SNARK gene was localized to human chromosome 1q32 by fluorescent in situ hybridization. SNARK was translated in vitro to yield a single protein band of approx. 76kDa; Western analysis of transfected baby hamster kidney (BHK) cells detected two SNARK-immunoreactive bands of approx. 76-80kDa. SNARK was capable of autophosphorylation in vitro; immunoprecipitated SNARK exhibited phosphotransferase activity with the synthetic peptide substrate HMRSAMSGLHLVKRR (SAMS) as a kinase substrate. SNARK activity was significantly increased by AMP and 5-amino-4-imidazolecarboxamide riboside (AICAriboside) in rat keratinocyte cells, implying that SNARK might be activated by an AMPK kinase-dependent pathway. Furthermore, glucose deprivation increased SNARK activity 3-fold in BHK fibroblasts. These findings identify SNARK as a glucose- and AICAriboside-regulated member of the AMPK-related gene family that represents a new candidate mediator of the cellular response to metabolic stress.

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Analysis of Htra Gene from Zebrafish (Danio Rerio)

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.7554 ◽

2015 ◽

Vol 10 (2) ◽

Author(s):

M. Murwantoko ◽

Chio Oka ◽

Masashi Kawaichi

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Danio Rerio ◽

Serine Proteases ◽

Catalytic Domain ◽

Fruit Fly ◽

Wide Range ◽

Igf Binding ◽

Bacterial Homolog

HtrA which is characterized by the combination of a trypsin-like catalytic domain with at least one C-terminalPDZ domain is a highly conserved family of serine proteases found in a wide range of organisms. However theidentified HtrA family numbers varies among spesies, for example the number of mammalian, Eschericia coli,fruit fly-HtrA family are 4, 3 and 1 gene respectively. One gene is predicted exist in zebrafish. Since no completeinformation available on zebrafish HtrA, in this paper zebrafish HtrA (zHtrA) gene was analyzed. The zHtrA isbelonged to HtrA1 member and predicted encodes 478 amino acids with a signal peptide, a IGF binding domain,a Kazal-type inhibitor domain in the up stream of HtrA-bacterial homolog. At the amino acid sequence the zHtrA1showed the 69%, 69%, 68%, 54% and 54% with the rat HtrA1, mouse HtrA1, human HtrA1, human HtrA3 andmouse HtrA4 respectively. The zHtrA1 is firstly expressed at 60 hpf and mainly in the vertebral rudiments in thetail region.

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Classification of Nonenzymatic Homologues of Protein Kinases

Comparative and Functional Genomics ◽

10.1155/2009/365637 ◽

2009 ◽

Vol 2009 ◽

pp. 1-17 ◽

Cited By ~ 5

Author(s):

K. Anamika ◽

K. R. Abhinandan ◽

K. Deshmukh ◽

N. Srinivasan

Keyword(s):

Protein Kinase ◽

Protein Kinases ◽

Protein Interaction ◽

Transmembrane Domain ◽

Homo Sapiens ◽

Interaction Domain ◽

Protein Protein Interaction ◽

Domain 3 ◽

Eukaryotic Organisms ◽

And Function

Protein Kinase-Like Non-kinases (PKLNKs), which are closely related to protein kinases, lack the crucial catalytic aspartate in the catalytic loop, and hence cannot function as protein kinase, have been analysed. Using various sensitive sequence analysis methods, we have recognized 82 PKLNKs from four higher eukaryotic organisms, namely,Homo sapiens,Mus musculus,Rattus norvegicus, andDrosophila melanogaster. On the basis of their domain combination and function, PKLNKs have been classified mainly into four categories: (1) Ligand binding PKLNKs, (2) PKLNKs with extracellular protein-protein interaction domain, (3) PKLNKs involved in dimerization, and (4) PKLNKs with cytoplasmic protein-protein interaction module. While members of the first two classes of PKLNKs have transmembrane domain tethered to the PKLNK domain, members of the other two classes of PKLNKs are cytoplasmic in nature. The current classification scheme hopes to provide a convenient framework to classify the PKLNKs from other eukaryotes which would be helpful in deciphering their roles in cellular processes.

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Diffusion rather than intraflagellar transport likely provides most of the tubulin required for axonemal assembly in Chlamydomonas

Journal of Cell Science ◽

10.1242/jcs.249805 ◽

2020 ◽

Vol 133 (17) ◽

pp. jcs249805 ◽

Cited By ~ 1

Author(s):

Julie Craft Van De Weghe ◽

J. Aaron Harris ◽

Tomohiro Kubo ◽

George B. Witman ◽

Karl F. Lechtreck

Keyword(s):

Binding Site ◽

Critical Concentration ◽

Intraflagellar Transport ◽

Microtubule Assembly ◽

Strong Reduction ◽

Tubulin Binding ◽

Respective Contribution ◽

Β Tubulin

ABSTRACTTubulin enters the cilium by diffusion and motor-based intraflagellar transport (IFT). However, the respective contribution of each route in providing tubulin for axonemal assembly remains unknown. Using Chlamydomonas, we attenuated IFT-based tubulin transport of GFP–β-tubulin by altering the IFT74N–IFT81N tubulin-binding module and the C-terminal E-hook of tubulin. E-hook-deficient GFP–β-tubulin was incorporated into the axonemal microtubules, but its transport frequency by IFT was reduced by ∼90% in control cells and essentially abolished when the tubulin-binding site of IFT81 was incapacitated. Despite the strong reduction in IFT, the proportion of E-hook-deficient GFP–β-tubulin in the axoneme was only moderately reduced. In vivo imaging showed more GFP–β-tubulin particles entering cilia by diffusion than by IFT. Extrapolated to endogenous tubulin, the data indicate that diffusion provides most of the tubulin required for axonemal assembly. We propose that IFT of tubulin is nevertheless needed for ciliogenesis, because it augments the tubulin pool supplied to the ciliary tip by diffusion, thus ensuring that free tubulin there is maintained at the critical concentration for plus-end microtubule assembly during rapid ciliary growth.

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Sequence, expression and localization of calmodulin-domain protein kinases inEimeria tenellaandEimeria maxima

Parasitology ◽

10.1017/s0031182000081506 ◽

1996 ◽

Vol 113 (5) ◽

pp. 439-448 ◽

Cited By ~ 21

Author(s):

P. P. J. Dunn ◽

J. M. Bumstead ◽

F. M. Tomley

Keyword(s):

Host Cell ◽

Protein Kinases ◽

Catalytic Domain ◽

Sequence Data ◽

Host Cells ◽

Cdna Clones ◽

Host Cell Membrane ◽

Amino Terminal ◽

Domain Protein

SUMMARYWe have isolated and sequenced cDNA clones fromEimeria tenellaandEimeria maximawhich encode proteins that share homology with a recently described family of calmodulin-domain protein kinases. The primary sequence data show that each of the protein kinases can be divided into 2 main functional domains – an amino-terminal catalytic domain typical of serine/threonine protein kinases and a carboxy-terminal domain homologous to calmodulin, which is capable of binding calcium ions at 4 ‘EF-hand’ motifs. Expression of theE. tenellacalmodulin-domain protein kinase (EtCDPK) increased towards the end of oocyst sporulation, as judged by Northern and Western blotting, and indirect immunofluorescent antibody labelling showed that within a few minutes of adding sporozoites to target host cells inin vitroculture EtCDPK was found to be specifically associated with a filament-like structure that converges at the apical end of the parasite. Once the parasite entered the host cell EtCDPK appeared to be left on the host cell membrane at the point of entry, indicating a brief yet specific role for this molecule in the invasion of host cells byE. tenella.

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The Roles of β-Tubulin Mutations and Isotype Expression in Acquired Drug Resistance

Cancer Informatics ◽

10.1177/117693510700300028 ◽

2007 ◽

Vol 3 ◽

pp. 117693510700300 ◽

Cited By ~ 35

Author(s):

J. Torin Huzil ◽

Ke Chen ◽

Lukasz Kurgan ◽

Jack A. Tuszynski

Keyword(s):

Drug Resistance ◽

Rational Design ◽

Resistant Cell ◽

Microtubule Assembly ◽

Acquired Drug Resistance ◽

Tubulin Isotypes ◽

Microtubule Disruption ◽

Tubulin Isotype ◽

Β Tubulin ◽

Isotype Expression

The antitumor drug paclitaxel stabilizes microtubules and reduces their dynamicity, promoting mitotic arrest and eventually apoptosis. Upon assembly of the α/β-tubulin heterodimer, GTP becomes bound to both the α and β-tubulin monomers. During microtubule assembly, the GTP bound to β-tubulin is hydrolyzed to GDP, eventually reaching steady-state equilibrium between free tubulin dimers and those polymerized into microtubules. Tubulin-binding drugs such as paclitaxel interact with β-tubulin, resulting in the disruption of this equilibrium. In spite of several crystal structures of tubulin, there is little biochemical insight into the mechanism by which anti-tubulin drugs target microtubules and alter their normal behavior. The mechanism of drug action is further complicated, as the description of altered β-tubulin isotype expression and/or mutations in tubulin genes may lead to drug resistance as has been described in the literature. Because of the relationship between β-tubulin isotype expression and mutations within β-tubulin, both leading to resistance, we examined the properties of altered residues within the taxane, colchicine and Vinca binding sites. The amount of data now available, allows us to investigate common patterns that lead to microtubule disruption and may provide a guide to the rational design of novel compounds that can inhibit microtubule dynamics for specific tubulin isotypes or, indeed resistant cell lines. Because of the vast amount of data published to date, we will only provide a broad overview of the mutational results and how these correlate with differences between tubulin isotypes. We also note that clinical studies describe a number of predictive factors for the response to anti-tubulin drugs and attempt to develop an understanding of the features within tubulin that may help explain how they may affect both microtubule assembly and stability.

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