scholarly journals The structure of the Myo4p globular tail and its function in ASH1 mRNA localization

2010 ◽  
Vol 189 (3) ◽  
pp. 497-510 ◽  
Author(s):  
Alexander Heuck ◽  
Ingrid Fetka ◽  
Daniel N. Brewer ◽  
Daniela Hüls ◽  
Mary Munson ◽  
...  
Keyword(s):  
Mrna Localization ◽  
Surface Patch ◽  
X Ray ◽  
Membrane Tethering ◽  
Dividing Cells ◽  
Dependent Transport ◽  
Type V ◽  
Ash1 Mrna

Type V myosin (MyoV)–dependent transport of cargo is an essential process in eukaryotes. Studies on yeast and vertebrate MyoV showed that their globular tails mediate binding to the cargo complexes. In Saccharomyces cerevisiae, the MyoV motor Myo4p interacts with She3p to localize asymmetric synthesis of HO 1 (ASH1) mRNA into the bud of dividing cells. A recent study showed that localization of GFP-MS2–tethered ASH1 particles does not require the Myo4p globular tail, challenging the supposed role of this domain. We assessed ASH1 mRNA and Myo4p distribution more directly and found that their localization is impaired in cells expressing globular tail–lacking Myo4p. In vitro studies further show that the globular tail together with a more N-terminal linker region is required for efficient She3p binding. We also determined the x-ray structure of the Myo4p globular tail and identify a conserved surface patch important for She3p binding. The structure shows pronounced similarities to membrane-tethering complexes and indicates that Myo4p may not undergo auto-inhibition of its motor domain.

The role of silicon in forages: A quantitative X-Ray study

1987 ◽  
Vol 45 ◽  
pp. 416-417
Author(s):  
Janet H. Woodward ◽  
D. E. Akin
Keyword(s):  
Sodium Acetate ◽  
Dry Matter ◽  
Acetate Buffer ◽  
Plant Tissues ◽  
Sodium Acetate Buffer ◽  
X Ray ◽  
Dry Matter Digestibility ◽  
Percent Composition

Silicon (Si) is distributed throughout plant tissues, but its role in forages has not been clarified. Although Si has been suggested as an antiquality factor which limits the digestibility of structural carbohydrates, other research indicates that its presence in plants does not affect digestibility. We employed x-ray microanalysis to evaluate Si as an antiquality factor at specific sites of two cultivars of bermuda grass (Cynodon dactvlon (L.) Pers.). “Coastal” and “Tifton-78” were chosen for this study because previous work in our lab has shown that, although these two grasses are similar ultrastructurally, they differ in in vitro dry matter digestibility and in percent composition of Si.Two millimeter leaf sections of Tifton-7 8 (Tift-7 8) and Coastal (CBG) were incubated for 72 hr in 2.5% (w/v) cellulase in 0.05 M sodium acetate buffer, pH 5.0. For controls, sections were incubated in the sodium acetate buffer or were not treated.

Effects of dietary alteration of bicarbonate and magnesium on rat bone

1986 ◽  
Vol 250 (2) ◽  
pp. F302-F307 ◽  
Author(s):  
J. M. Burnell ◽  
C. Liu ◽  
A. G. Miller ◽  
E. Teubner
Keyword(s):  
Crystal Structure ◽  
Bone Formation ◽  
X Ray Diffraction ◽  
X Ray ◽  
Growing Rats ◽  
Final Composition ◽  
Rat Bone

To study the effects of bicarbonate and magnesium on bone, mild acidosis and/or hypermagnesemia were produced in growing rats by feeding ammonium chloride and/or magnesium sulfate. Bone composition, quantitative histomorphometry, and mineral x-ray diffraction (XRD) characteristics were measured after 6 wk of treatment. The results demonstrated that both acidosis (decreased HCO3) and hypermagnesemia inhibited periosteal bone formation, and, when combined, results were summative; and the previously observed in vitro role of HCO3- and Mg2+ as inhibitors of crystal growth were confirmed in vivo. XRD measurements demonstrated that decreased plasma HCO3 resulted in larger crystals and increased Mg resulted in smaller crystals. However, the combined XRD effects of acidosis and hypermagnesemia resembled acidosis alone. It is postulated that the final composition and crystal structure of bone are strongly influenced by HCO3- and Mg2+, and the effects are mediated by the combined influence on both osteoblastic bone formation and the growth of hydroxyapatite.

scholarly journals Promising Role of Calcium Hypochlorite as a Disinfectant: An in vitro Evaluation Regarding its Effect on Type V Dental Stone

2012 ◽  
Vol 13 (6) ◽  
pp. 856-866
Author(s):  
Prasanta Kumar Swain ◽  
Sharanbasappa C Nagaral ◽  
Pawan Kumar Kamalapurker ◽  
Ravishankar Damineni
Keyword(s):  
Tensile Strength ◽  
Bacillus Subtilis ◽  
Deleterious Effect ◽  
Strength Testing ◽  
Calcium Hypochlorite ◽  
Resistant Species ◽  
Type V ◽  
Mixing Water

ABSTRACT Aim The current study has been chosen to evaluate the efficacy of calcium hypochlorite as a disinfecting additive for the gypsum products and its effect on compressive and tensile strength of the set material. It is hypothesized that, the addition of calcium hypochlorite to type V dental stone in sufficient quantity to disinfect the material would have no deleterious effect on compressive or tensile strength. Materials and methods Total of 160 samples made up of type V dental stone were divided broadly into two groups of 80 samples each for the sake of compressive and tensile strength testing in dry and wet conditions: Out of each group, 10 samples without addition of any disinfectant (0% calcium hypochlorite) was compared with other group of 30 samples after adding disinfectant, i.e. each subgroup containing 10 samples each (0.5, 1.0 and 1.5% calcium hypochlorite). Conclusion Within limitations of this in vitro study it is assumed to prepare type V dental stone that contains a disinfectant, has adequate compressive strength and tensile strength, and can significantly act against a resistant species like Bacillus subtilis. Clinical significance When calcium hypochlorite was added to dental stone, extra mixing water was required to produce a material of nearly same pouring consistency. The samples, which were put to microbiological tests, showed effective action of disinfectant on Bacillus subtilis. No deleterious effect on compressive or tensile strength could be found after putting the selected samples with calcium hypochlorite. How to cite this article Swain PK, Nagaral SC, Kamalapurker PK, Damineni R. Promising Role of Calcium Hypochlorite as a Disinfectant: An in vitro Evaluation Regarding its Effect on Type V Dental Stone. J Contemp Dent Pract 2012;13(6):856-866.

Coordinate expression of secretory phospholipase A2 and cyclooxygenase-2 in activated human keratinocytes

2000 ◽  
Vol 278 (4) ◽  
pp. C822-C833 ◽  
Author(s):  
Krystyna E. Rys-Sikora ◽  
Raymond L. Konger ◽  
John W. Schoggins ◽  
Rama Malaviya ◽  
Alice P. Pentland
Keyword(s):  
Wound Repair ◽  
Human Keratinocytes ◽  
Plating Density ◽  
Coordinate Expression ◽  
Cox 2 ◽  
Type V ◽  
In Vitro Wounding

PGE2 levels are altered in human epidermis after in vivo wounding; however, mechanisms modulating PGE2 production in activated keratinocytes are unclear. In previous studies, we showed that PGE2 is a growth-promoting autacoid in human primary keratinocyte cultures, and its production is modulated by plating density, suggesting that regulated PGE2 synthesis is an important component of wound healing. Here, we examine the role of phospholipase A2(PLA2) and cyclooxygenase (COX) enzymes in modulation of PGE2 production. We report that the increased PGE2 production that occurs in keratinocytes grown in nonconfluent conditions is also observed after in vitro wounding, indicating that similar mechanisms are involved. This increase was associated with coordinate upregulation of both COX-2 and secretory PLA2 (sPLA2) proteins. Increased sPLA2 activity was also observed. By RT-PCR, we identified the presence of type IIA and type V sPLA2, along with the M-type sPLA2 receptor. Thus the coordinate expression of sPLA2 and COX-2 may be responsible for the increased prostaglandin synthesis in activated keratinocytes during wound repair.

scholarly journals Bacterial dynamin-like protein DynA mediates lipid and content mixing and shows phospholipid specificity

2018 ◽  
Author(s):  
Lijun Guo ◽  
Marc Bramkamp
Keyword(s):  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Membrane Stress ◽  
Gtp Hydrolysis ◽  
Cellular Processes ◽  
Organelle Division ◽  
Membrane Tethering ◽  
Hydrolysis Of

ABSTRACTThe dynamins family of GTPases is involved in key cellular processes in eukaryotes, including vesicle trafficking and organelle division. The GTP hydrolysis cycle of dynamin translates to a conformational change in the protein structure, which forces the underlying lipid layer into an energetically unstable conformation that promotes membrane rearrangements. Many bacterial genomes encode dynamin-like proteins, but the biological function of these proteins has remained largely enigmatic. In recent years, our group has reported that the dynamin-like protein DynA from Bacillus subtilis mediates nucleotide-independent membrane tethering in vitro and contributes to the innate immunity of bacteria against membrane stress and phage infection. However, so far the mechanism of membrane stress response and the role of GTP hydrolysis remain unclear. Here, we employed content mixing and lipid mixing assays in reconstituted systems to study if the dynamin-like protein DynA from B. subtilis induces membrane full fusion, and further test the possibility that GTP hydrolysis of DynA may act on the fusion-through-hemifusion pathway. Our results based on fluorescence resonance energy transfer (FRET) indicated that DynA could induce aqueous content mixing even in absence of GTP. Moreover, DynA-induced membrane fusion in vitro is a thermo-promoted slow response. Surprisingly, digestion of protein mediated an instantl rise of content exchange, supporting the assumption that disassembly of DynA is the fundamental power for fusion-through-hemifusion.

scholarly journals Chronology of Formation of Solar Radio Burst Types III and V Associated with Solar Flare Phenomenon on 19th September 2011

2013 ◽  
Vol 24 ◽  
pp. 32-42
Author(s):  
Zety Sharizat Hamidi ◽  
N.N.M. Shariff
Keyword(s):  
Active Region ◽  
Solar Flare ◽  
Solar Radio ◽  
Active Regions ◽  
Radio Flux ◽  
X Ray ◽  
Type Iii ◽  
Solar Bursts ◽  
Type V

The formation of two different solar bursts, type III and V in one solar flare event is presented. Both bursts are found on 19th September 2011 associated with C-class flares on active region 1295. From the observation, we believed that the mechanism of evolution the bursts play an important role in the event. It is found that type V burst appeared in five minutes after type III. There are a few active regions on the solar disk but most are magnetically simple and have remained rather quiet. An interpretation of this new result depends critically on the number of sunspots and the role of active region 1295. Sunspot number is increased up to 144 with seven sunspots can be observed. During that event, the speed of solar wind exceeds 433.8 km/second with 2.0 g/cm3 density of protons in the solar corona. Currently, radio flux is also high up to 150 SFU. The solar flare type C6 is continuously being observed in the X-ray region for 24 hours since 1541 UT and a maximum C1 is detected on 1847 UT. Although the sources of both bursts are same, the direction and ejection explode differently. It is believed that the ejection of particles in a type III burst is higher than solar burst type V.

scholarly journals She3p Possesses a Novel Activity Required for ASH1 mRNA Localization in Saccharomyces cerevisiae

2009 ◽  
Vol 8 (7) ◽  
pp. 1072-1083 ◽  
Author(s):  
Sharon M. Landers ◽  
Michelle R. Gallas ◽  
Jaime Little ◽  
Roy M. Long
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Rna Binding ◽  
Adaptor Protein ◽  
Mrna Localization ◽  
Rna Localization ◽  
The Novel ◽  
Specific Proteins ◽  
Type V ◽  
Localization Elements ◽  
Ash1 Mrna

ABSTRACT Intracellular and intercellular polarity requires that specific proteins be sorted to discreet locations within and between cells. One mechanism for sorting proteins is through RNA localization. In Saccharomyces cerevisiae, ASH1 mRNA localizes to the distal tip of the bud, resulting in the asymmetric sorting of the transcriptional repressor Ash1p. ASH1 mRNA localization requires four cis-acting localization elements and the trans-acting factors Myo4p, She3p, and She2p. Myo4p is a type V myosin motor that functions to directly transport ASH1 mRNA to the bud. She2p is an RNA-binding protein that directly interacts with the ASH1 mRNA cis-acting elements. Currently, the role for She3p in ASH1 mRNA localization is as an adaptor protein, since it can simultaneously associate with Myo4p and She2p. Here, we present data for two novel mutants of She3p, S348E and the double mutant S343E S361E, that are defective for ASH1 mRNA localization, and yet both of these mutants retain the ability to associate with Myo4p and She2p. These observations suggest that She3p possesses a novel activity required for ASH1 mRNA localization, and our data imply that this function is related to the ability of She3p to associate with ASH1 mRNA. Interestingly, we determined that She3p is phosphorylated, and global mass spectrometry approaches have determined that Ser 343, 348, and 361 are sites of phosphorylation, suggesting that the novel function for She3p could be negatively regulated by phosphorylation. The present study reveals that the current accepted model for ASH1 mRNA localization does not fully account for the function of She3p in ASH1 mRNA localization.

scholarly journals Dissecting the roles of Haspin and VRK1 in Histone H3 threonine-3 phosphorylation during mitosis

2021 ◽  
Author(s):  
Tyrell N Cartwright ◽  
Rebecca J Harris ◽  
Stephanie K Meyer ◽  
Nikolaus A. Watson ◽  
Cheryl Tan ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Neuromuscular Disease ◽  
Kinase Activity ◽  
Histone H3 ◽  
Direct Role ◽  
Dividing Cells ◽  
Histone Kinase ◽  
Mitosis And Meiosis

Protein kinases that phosphorylate histones are ideally-placed to influence the behavior of chromosomes during cell division. Indeed, a number of conserved histone phosphorylation events occur prominently during mitosis and meiosis in most eukaryotes, including on histone H3 at threonine-3 (H3T3ph). At least two kinases, Haspin and VRK1 (NHK-1/ballchen in Drosophila), have been proposed to carry out this modification. Phosphorylation of H3 by Haspin has defined roles in mitosis, but the significance of VRK1 activity towards histones in dividing cells has been unclear. Here, using in vitro kinase assays, KiPIK screening, RNA interference, and CRISPR/Cas9 approaches, we were unable to substantiate a direct role for VRK1, or its homologue VRK2, in the phosphorylation of threonine-3 or serine-10 of Histone H3 in mitosis, although loss of VRK1 did slow cell proliferation. We conclude that the role of VRK1, and its more recently identified association with neuromuscular disease in humans, is unlikely to involve mitotic histone kinase activity. In contrast, Haspin is required to generate H3T3ph during mitosis.

scholarly journals Exoribonuclease and Endoribonuclease Activities of RNase BN/RNase Z both Function in Vivo

2012 ◽  
Vol 287 (42) ◽  
pp. 35747-35755 ◽  
Author(s):  
Tanmay Dutta ◽  
Arun Malhotra ◽  
Murray P. Deutscher
Keyword(s):  
Potential Role ◽  
Wild Type ◽  
X Ray ◽  
E Coli ◽  
Phage T4 ◽  
A Cell ◽  
Rnase Z

Escherichia coli RNase BN, a member of the RNase Z family of endoribonucleases, differs from other family members in that it also can act as an exoribonuclease in vitro. Here, we examine whether this activity of RNase BN also functions in vivo. Comparison of the x-ray structure of RNase BN with that of Bacillus subtilis RNase Z, which lacks exoribonuclease activity, revealed that RNase BN has a narrower and more rigid channel downstream of the catalytic site. We hypothesized that this difference in the putative RNA exit channel might be responsible for the acquisition of exoribonuclease activity by RNase BN. Accordingly, we generated several mutant RNase BN proteins in which residues within a loop in this channel were converted to the corresponding residues present in B. subtilis RNase Z, thus widening the channel and increasing its flexibility. The resulting mutant RNase BN proteins had reduced or were essentially devoid of exoribonuclease activity in vitro. Substitution of one mutant rbn gene (P142G) for wild type rbn in the E. coli chromosome revealed that the exoribonuclease activity of RNase BN is not required for maturation of phage T4 tRNA precursors, a known specific function of this RNase. On the other hand, removal of the exoribonuclease activity of RNase BN in a cell lacking other processing RNases leads to slower growth and affects maturation of multiple tRNA precursors. These findings help explain how RNase BN can act as both an exo- and an endoribonuclease and also demonstrate that its exoribonuclease activity is capable of functioning in vivo, thus widening the potential role of this enzyme in E. coli.

scholarly journals An Exclusively Nuclear RNA-Binding Protein Affects Asymmetric Localization of ASH1 mRNA and Ash1p in Yeast

2001 ◽  
Vol 153 (2) ◽  
pp. 307-318 ◽  
Author(s):  
Roy M. Long ◽  
Wei Gu ◽  
Xiuhua Meng ◽  
Graydon Gonsalvez ◽  
Robert H. Singer ◽  
...  
Keyword(s):  
Rna Binding ◽  
Rna Binding Proteins ◽  
Mrna Localization ◽  
Yeast Cells ◽  
Rna Structures ◽  
Nuclear Factors ◽  
Mother Cells ◽  
Ash1 Mrna

The localization of ASH1 mRNA to the distal tip of budding yeast cells is essential for the proper regulation of mating type switching in Saccharomyces cerevisiae. A localization element that is predominantly in the 3′-untranslated region (UTR) can direct this mRNA to the bud. Using this element in the three-hybrid in vivo RNA-binding assay, we identified a protein, Loc1p, that binds in vitro directly to the wild-type ASH1 3′-UTR RNA, but not to a mutant RNA incapable of localizing to the bud nor to several other mRNAs. LOC1 codes for a novel protein that recognizes double-stranded RNA structures and is required for efficient localization of ASH1 mRNA. Accordingly, Ash1p gets symmetrically distributed between daughter and mother cells in a loc1 strain. Surprisingly, Loc1p was found to be strictly nuclear, unlike other known RNA-binding proteins involved in mRNA localization which shuttle between the nucleus and the cytoplasm. We propose that efficient cytoplasmic ASH1 mRNA localization requires a previous interaction with specific nuclear factors.

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