117,829 research outputs found
Exploring the diversity of SPRY/B30.2-mediated interactions
No full textThe SPla/Ryanodine receptor (SPRY)/B30.2 domain is one of the most common folds in higher eukaryotes. The human genome encodes 103 SPRY/B30.2 domains, several of which are involved in the immune response. Approximately 45% of human SPRY/B30.2-containing proteins are E3 ligases. The role and function of the majority of SPRY/B30.2 domains are still poorly understood, however, in several cases mutations in this domain have been linked to congenital disorders. The recent characterization of SPRY/B30.2-mediated protein interactions has provided evidence for a role of this domain as an adaptor module to assemble macromolecular complexes, analogous to Src homology (SH)2, SH3, and WW domains. However, functional and structural evidence suggests that SPRY/B30.2 is a more versatile fold, allowing a wide range of binding modes
L-M
No full textSeries 2941 | Governor (1909-1917: Spry) | Joseph Hillstrom Correspondence | L-MAs chief executive officer of the state, the governor has the authority to commute the sentences of those convicted of crimes. The conviction and execution of Joseph Hillstrom (Joe Hill) became one of the most controversial criminal cases in Utah history; it generated national and international interest. Correspondence is in the form of letters, postcards, and telegrams
L: July-Aug. 1914; May-Sept. 1915
No full textSeries 2941 | Governor (1909-1917: Spry) | Joseph Hillstrom Correspondence | L: July-Aug. 1914; May-Sept. 1915As chief executive officer of the state, the governor has the authority to commute the sentences of those convicted of crimes. The conviction and execution of Joseph Hillstrom (Joe Hill) became one of the most controversial criminal cases in Utah history; it generated national and international interest. Correspondence is in the form of letters, postcards, and telegrams
The Role of the SPRY domain in the SPRY domain-containing SOCS box proteins
No full text© 2005 Seth L. Masters.There are four mammalian SSB proteins (SSB-1 to -4), and these are characterized by a C-terminal SOCS box and central SPRY domain. The C-terminal SOCS box was first observed in proteins that were found to act as Suppressors of Cytokine Signalling and function by virtue of their SH2 domain. Other families containing the SOCS box motif were defined by the domains N-terminal to this, such as the ASBs (Ankyrin repeats), WSBs (WD40 repeats) and of course the SSBs (SPRY domains). This thesis describes a very broad investigation of the SSBs, a protein family about which very little was known. To begin with, functional investigation into the evolution of this family and analysis of murine SSB expression patterns was performed. This highlighted that the family was highly conserved and had differential expression in the mouse, suggestive of important, unique functional roles for the individual family members. The majority of work in the thesis then proceeds in three directions; (i) analysis of the SSB proteins in vivo, with genetic deletion of SSB-2 in the mouse, (ii) biochemically, with analysis of SSB binding partners, and (iii) structurally, with functional analysis of the structure of SSB-2
The Effect of New Jersey Lottery Promotions on Consumer Demand and State Profits
No full textWe estimate elasticities of demand for New Jersey’s Pick 3 and Pick 4 midday/evening numbers games by exploiting random price variation generated by episodic promotions for each game. These Pick 3 Green Ball and Pick 4 Red Ball promotions lower the price of a lottery ticket for an evening numbers game by increasing prize payments during the 28-day promotion periods. The own-price elasticities of demand for the evening Pick 3 and Pick 4 games are both approximately -0.5. During the promotions, the loss in profit margins outweighs the gain in sales because of this inelastic demand. However, the combined effects of lower evening Pick 3 profits and increased sales of complementary products boost lottery profits by 840,000 during the 28 days of the Green Ball promotion, while the combined effects of lower evening Pick 4 profits and reduced sales of substitute products decrease lottery profits by 3.61 million during the 28 days of the Red Ball promotion. If higher sales after the promotion are included, the total increase in profits potentially reaches $14.48 million under the Green Ball game, while the Red Ball promotion loses money for the lottery even considering its positive lagged effect.Peer reviewedThis is a post-peer-review, pre-copyedit version of an article published in Eastern Economic Journal. The definitive publisher-authenticated version (Combs, Kathryn L., Jocelyn Elise Crowley, and John A. Spry, "The Effect of New Jersey Lottery Promotions on Consumer Demand and State Profits," Eastern Economic Journal (2014) 40, 326–348. doi:10.1057/eej.2013.39) is available online at: http://dx.doi.org/10.1057/eej.2013.39
Unraveling the mechanisms of PAMless DNA interrogation by SpRY-Cas9
No full textCRISPR-Cas9 is a powerful tool for genome editing, but the strict requirement for an NGG protospacer-adjacent motif (PAM) sequence immediately next to the DNA target limits the number of editable genes. Recently developed Cas9 variants have been engineered with relaxed PAM requirements, including SpG-Cas9 (SpG) and the nearly PAM-less SpRY-Cas9 (SpRY). However, the molecular mechanisms of how SpRY recognizes all potential PAM sequences remains unclear. Here, we combine structural and biochemical approaches to determine how SpRY interrogates DNA and recognizes target sites. Divergent PAM sequences can be accommodated through conformational flexibility within the PAM-interacting region, which facilitates tight binding to off-target DNA sequences. Nuclease activation occurs ~1000-fold slower than for Streptococcus pyogenes Cas9, enabling us to directly visualize multiple on-pathway intermediate states. Experiments with SpG position it as an intermediate enzyme between Cas9 and SpRY. Our findings shed light on the molecular mechanisms of PAMless genome editing
The SPRY domain of TRIM25 associates with ZAP.
No full text(A) 293T cells were infected with the SINV strain Toto1101 (MOI = 10), and lysates were harvested at 0, 6, 12 and 24 h p.i. for co-immunoprecipitation with an anti-TRIM25 antibody and immunoblotting. The data is representative of 2 independent experiments. (B) Human ZAP short (S) and long (L) isoforms are shown, with the CCCH fingers, TIPARP, WWE and PARP-like domains indicated. Light green shading indicates sequences shared by the two isoforms whereas the hatched region containing the PARP-like domain is unique to the L isoform. WCL of ZC3HAV1-knockout 293T cells transfected with V5-tagged TRIM25 and/or ZAPS or ZAPL were used for co-immunoprecipitation with an anti-NZAP antibody and immunoblotting. (C) Full-length (FL) TRIM25 is shown, with the RING, B box, CCD, and SPRY domains indicated. WCL of ZC3HAV1-knockout 293T cells transfected with V5-tagged FL or truncated TRIM25 (RING, B box/CCD, SPRY only) and ZAPL were used for co-immunoprecipitation with an anti-NZAP antibody and immunoblotting. Different amounts of TRIM25 domain-expressing constructs were used for transfection in order to achieve similar RING, B box/CCD, and SPRY expression. The asterisk indicates a non-specific band. (B and C) The data is representative of 3 independent experiments.</p
Respon Pertumbuhan dan Produksi Kacang Ijo (Vigna radiata L.) Dengan Pemberian Dosis Pupuk Spry Up dan SP-36
No full textMung bean (Vigna radiata L.) is one of the important food commodities in Indonesia, but its productivity is still low. One of the efforts to increase the productivity of mung beans is with balanced fertilization according to plant needs. The purpose of this study was to determine the interaction of dosage of spry up liquid fertilizer and SP-36 to produce the best growth and highest production. This research was a factorial experiment using a completely randomized design with two treatment factors and three replications. The first factor was the dose of liquid fertilizer spry up, with three levels, namely 1; 1.5; and 2 ml/plant. The second factor was fertilizer SP-36, with three levels, namely 5; 7; and 10 gr/plant. Observation data were analyzed with analysis of variance and continued with the LSD 5% if the results were significant. The results of the experiment showed that there was significant difference in the parameters of plant height, number of leaves, leaf area, and the amount of mung bean production at each increase in the treatment. The combination of treatment at the of liquid fertilizer spry up of 2 ml/plant and SP-36 10 gr/plant (S3P3) showed the largest yield, namely plant height 43.110 cm, the number of leaves 31 strands with a production of 97.985 gr/plant. Kacang hijau (Vigna radiata L.) merupakan salah satu komoditas pangan penting di Indonesia, namun produktivitasnya masih rendah. Salah satu usaha peningkatan produktivitas kacang hijau dengan pemupukan yang berimbang sesuai kebutuhan tanaman. Tujuan dari penelitian ini adalah mengetahui interaksi dosis pupuk cair spry up dan SP-36 untuk menghasilkan pertumbuhan terbaik dan produksi paling tinggi. Penelitian ini merupakan percobaan faktorial menggunakan Rancangan Acak Lengkap dengan dua faktor perlakuan dan tiga kali ulangan. Faktor pertama adalah dosis pupuk cair spry up dengan tiga taraf yaitu 1; 1.5; dan 2 ml/tanaman. Faktor kedua adalah pupuk SP-36 dengan tiga taraf yaitu 5; 7; dan 10 gr/tanaman. Data hasil pengamatan dianalisis dengan analisis varians dan dilanjutkan uji BNT 5% apabila hasil signifikan. Hasil percobaan menunjukkan bahwa terjadi interaksi yang berbeda nyata pada parameter tinggi tanaman, jumlah daun, luas daun dan jumlah produksi tanaman kacang hijau pada setiap kenaikan dosis perlakuan. Kombinasi perlakuan pada dosis pupuk cair spry up 2 ml/tanaman dan SP-36 10 gr/tanaman (S3P3) menunjukkan hasil terbesar yaitu tinggi tanaman 43,110 cm, jumlah daun 31 helai dengan produksi sebesar 97,985 gram/tanaman
New structural data reveal benleonardite to be a member of the pearceite-polybasite group
No full textNHM Repositor
PAM-interacting domain turn-helix 51 motifs can improve Cas9–SpRY activity
No full textCas9-SpRY is an engineered variant of the Streptococcus pyogenes Cas9 with relaxed PAM recognition, which can technically be utilized at any target in the genome but some targets are addressed with low efficiency. Here, we show that a previously unexplored motif at the turn and beginning of α-helix 51 (TH51) can be engineered to improve both nuclease and prime-editing activity of Cas9-SpRY. Interaction of the lysine-rich PID loop 2 (PL2) with the target DNA downstream of the PAM (post-PAM) mediates initiation of R-loop formation and subsequent cleavage yet it was unclear if other regions of the PID engage with post-PAM as well. To this end, the NAAN-PAM-targeting iSpyMac hybrid nuclease, which lacks all lysine residues in PL2, was compared with Cas9-SpRY at identical targets using molecular dynamics simulation and in cell culture models, uncovering four crucial post-PAM-interacting lysines in TH51 and TH53 of iSpyMac. Ectopic insertion of a lysine-rich PL2 into iSpyMac boosted its nuclease and prime-editing activities and, in turn, Cas9-SpRY benefited from certain lysine-rich TH51 motifs. Specifically, TH51 from an uncultured Abiotrophia Cas9 species boosted overall Cas9-SpRY activity. Together, this study demonstrates that engineering of post-PAM interacting motifs opens new avenues for the design of advanced CRISPR enzymes
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