排序方式: 共有149条查询结果,搜索用时 15 毫秒
31.
Nathanael R. Hauck Hisayo Yamane Ryutaro Tao Amy F. Iezzoni 《Sexual plant reproduction》2002,15(1):39-46
. Gametophytic self-incompatibility (GSI) typically "breaks down" due to polyploidy in many Solanaceous species, resulting in self-compatible (SC) tetraploid individuals. However, sour cherry (Prunus cerasus L.), a tetraploid species resulting from hybridization of the diploid sweet cherry (P. avium L.) and the tetraploid ground cherry (P. fruticosa Pall.), is an exception, consisting of both self-incompatible (SI) and SC individuals. Since sweet cherry exhibits GSI with 13 S-ribonucleases (S-RNases) identified as the stylar S-locus product, the objectives were to compare sweet and sour cherry S-allele function, S-RNase sequences and linkage map location as initial steps towards understanding the genetic basis of SI and SC in sour cherry. S-RNases from two sour cherry cultivars that were the parents of a linkage mapping population were cloned and sequenced. The sequences of two S-RNases were identical to those of sweet cherry S-RNases, whereas three other S-RNases had unique sequences. One of the S-RNases mapped to the Prunus linkage group 6, similar to its location in sweet cherry and almond, whereas two other S-RNases were linked to each other but were unlinked to any other markers. Interspecific crosses between sweet and sour cherry demonstrated that GSI exists in sour cherry and that the recognition of common S-alleles has been maintained in spite of polyploidization. It is hypothesized that self-compatibility in sour cherry is caused by the existence of non-functional S-RNases and pollen S-genes that may have arisen from natural mutations. 相似文献
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Nathanael Fickett Andres Gutierrez Mohit Verma Michael Pontif Anna Hale Collins Kimbeng Niranjan Baisakh 《Genomics》2019,111(6):1794-1801
Sugarcane is an economically important crop for both food and biofuel industries. Marker-assisted breeding in sugarcane is becoming a reality with the recent development and deployment of markers linked with disease resistance genes. Large linkage disequilibrium in sugarcane makes genome-wide association studies (GWAS) a better alternative to biparental mapping to identify markers associated with agronomic traits. GWAS was conducted on a Louisiana core collection to identify marker-trait associations (MTA) for 11 cane yield and sucrose traits using single nucleotide polymorphism (SNP) and insertion-deletion (Indel) markers. Significant (P < .05) MTAs were identified for all traits where the top ranked markers explained up to 15% of the total phenotypic variation. High correlations (0.732 to 0.999) were observed between sucrose traits and 56 markers were found consistent across multiple traits. These markers following validation in more diverse populations could be used in marker-assisted selection of clones in sugarcane breeding program in Louisiana and elsewhere. 相似文献
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Deng X Zhou W Weisberg E Wang J Zhang J Sasaki T Nelson E Griffin JD Jänne PA Gray NS 《Bioorganic & medicinal chemistry letters》2012,22(14):4579-4584
Here we describe the synthesis and characterization of a number of 3-amino-1H-indazol-6-yl-benzamides that were designed to target the 'DFG-out' conformation of the kinase activation loop. Several compounds such as 4 and 11 exhibit single-digit nanomolar EC(50)s against FLT3, c-Kit and the gatekeeper T674M mutant of PDGFRα. 相似文献
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Billecke N Raschzok N Rohn S Morgul MH Schwartlander R Mogl M Wollersheim S Schmitt KR Sauer IM 《Journal of biotechnology》2012,159(1-2):83-89
Cell morphology, proliferation and motility, as well as mono- and heterotypic cell-to-cell interactions, are of increasing interest for in vitro experiments. However, tightly controlling culture conditions whilst simultaneously monitoring the same set of cells is complicated. Moreover, video-microscopy of distinct cells or areas of cells over a prolonged period of time represents a technical challenge. The SlideObserver was designed for cinemicrography of cells in co-and monoculture. The core elements of the system are the SlideReactors, miniaturised hollow fibre-based bioreactors operated in closed perfusion loops. Within the SlideReactors, cells can be cultured under adaptable conditions as well as in direct- and indirect co-culture. The independent perfusion loops enable controlled variation of parameters such as medium, pH, and oxygenation. A combined automated microscope stage and camera set-up allows for micrograph acquisition of multiple user-defined regions of interest within the bioreactor units. For proof of concept, primary cells (HUVEC, human hepatocytes) and cell lines (HuH7, THP-1) were cultured under stable and varying culture conditions, as well as in mono- and co-culture. The operational system enabled non-stop imaging and automated control of process parameters as well as elective manipulation of either reactor. As opposed to non-perfused culture systems or comparable devices for cinemicrographic analysis, the SlideObserver allows simultaneous morphological monitoring of an entire culture of cells in multiple bioreactors. 相似文献
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Gametophytic self-incompatibility (GSI) is an outcrossing mechanism in flowering plants that is genetically controlled by 2 separate genes located at the highly polymorphic S-locus, termed S-haplotype. This study characterizes a pollen part mutant of the S(1)-haplotype present in sour cherry (Rosaceae, Prunus cerasus L.) that contributes to the loss of GSI. Inheritance of S-haplotypes from reciprocal interspecific crosses between the self-compatible sour cherry cultivar Ujfehértói Fürt?s carrying the mutated S(1)-haplotype (S(1)'S(4)S(d)S(null)) and the self-incompatible sweet cherry (Prunus avium L.) cultivars carrying the wild-type S(1)-haplotype revealed that the mutated S(1)-haplotype confers unilateral incompatibility with a functional pistil component and a nonfunctional pollen component. The altered sour cherry S(1)-haplotype pollen part mutant, termed S(1)', contains a 615-bp Ds-like element within the S(1)-haplotype-specific F-box protein gene (SFB(1)'). This insertion generates a premature in-frame stop codon that would result in a putative truncated SFB(1) containing only 75 of the 375 amino acids present in the wild-type SFB(1). S(1)' along with 2 other previously characterized Prunus S-haplotype mutants, S(f) and S(6m), illustrate that mobile element insertion is an evolutionary force contributing to the breakdown of GSI. 相似文献
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Yongmun Choi Markus A. Seeliger Shoghag B. Panjarian Hakjoong Kim Xianming Deng Taebo Sim Brian Couch Anthony J. Koleske Thomas E. Smithgall Nathanael S. Gray 《The Journal of biological chemistry》2009,284(42):29005-29014
Allosteric kinase inhibitors hold promise for revealing unique features of kinases that may not be apparent using conventional ATP-competitive inhibitors. Here we explore the activity of a previously reported allosteric inhibitor of BCR-Abl kinase, GNF-2, against two cellular isoforms of Abl tyrosine kinase: one that carries a myristate in the N terminus and the other that is deficient in N-myristoylation. Our results show that GNF-2 inhibits the kinase activity of non-myristoylated c-Abl more potently than that of myristoylated c-Abl by binding to the myristate-binding pocket in the C-lobe of the kinase domain. Unexpectedly, indirect immunofluorescence reveals a translocation of myristoylated c-Abl to the endoplasmic reticulum in GNF-2-treated cells, whereas GNF-2 has no detectable effect on the localization of non-myristoylated c-Abl. These results indicate that GNF-2 competes with the NH2-terminal myristate for binding to the c-Abl kinase myristate-binding pocket and that the exposed myristoyl group accounts for the localization to the endoplasmic reticulum. We also demonstrate that GNF-2 can inhibit enzymatic and cellular kinase activity of Arg, a kinase highly homologous to c-Abl, which is also likely to be regulated through intramolecular binding of an NH2-terminal myristate lipid. These results suggest that non-ATP-competitive inhibitors, such as GNF-2, can serve as chemical tools that can discriminate between c-Abl isoform-specific behaviors.The catalytic activity of a protein kinase can be modulated by binding of a ligand to a site distant from the active site, also referred to as the allosteric site (1). The ligand is referred to as an allosteric kinase inhibitor and induces a protein conformation that is not compatible with kinase activity. Allosteric inhibitors can potentially be exploited to elucidate kinase functions not discovered using ATP-competitive inhibitors, because they can exploit binding sites and regulatory mechanisms that are unique to a particular kinase.The c-Abl and Arg (Abl-related gene) proteins comprise the Abl family of non-receptor tyrosine kinases. Each family member has two isoforms: one that is myristoylated in the N terminus (1b or IV) and the other that is deficient in N-myristoylation due to an alternative splicing of the first exon (1a or I) (Fig. 1A). N-Myristoylation often serves as a mechanism for targeting proteins to cellular membranes. However, Abl family members localize to multiple subcellular compartments; whereas Arg is mostly found in the cytoplasm, c-Abl shuttles between the nucleus and the cytoplasm, where it localizes to the cytosol, endoplasmic reticulum, and mitochondria (2).Open in a separate windowFIGURE 1.A, domain structure of Abl family members (5). The numbers indicate amino acid residues in c-Abl 1b, and the recombinant protein constructs used in this study encompass amino acids 65–534, 83–534, and 248–531. B, ribbon representation of the c-Abl kinase NH2-terminal half residues, including the SH3, SH2, and kinase domains (Protein Data Bank code 1OPK) (7). The NH2-terminal cap (amino acids 2–79) is indicated by dotted lines (8). The myristate-binding site and ATP binding pocket are indicated by arrows. C, ribbon representation of an enlarged view of GNF-2 (colored gold) bound to the c-Abl myristate binding site. The location of Ala356 is indicated.The Abl family members share a high degree of sequence identity (∼90%) in the NH2-terminal half residues, including the SH3,2 SH2, and kinase domains (3). The kinase domain is followed by proline-rich motifs that serve as binding sites for SH3 domains. A range of proteins are reported to bind directly or indirectly to the SH3, SH2, and proline-rich domains of c-Abl and are implicated in the proper regulation of the kinase activities of Abl family members in the cytoplasm (4–6). In addition, as revealed by recent crystallographic analyses of inactive and assembled form of recombinant Abl, the kinase activity of c-Abl is modulated by the intrinsic binding of the N-myristoyl residue to a hydrophobic pocket in the C-lobe of the kinase domain, which induces conformational changes in the kinase domain and subsequently allows the SH3 and SH2 domains to pack against the kinase domain (7, 8). Altogether, these observations suggest that the kinase activities of Abl family members in normal cells are tightly regulated by both intra- and intermolecular interactions (2, 9). Disruption of these strong regulatory mechanisms results in deregulated kinase activity, as illustrated by the BCR-Abl and v-Abl oncoproteins.Recent years have seen great advances in pharmacological inhibition of deregulated c-Abl kinase activity. Among the small molecule inhibitors targeting BCR-Abl kinase are imatinib (STI-571; Gleevec), nilotinib (AMN 107), and dasatinib (BMS-354825) (10). These small molecules have been used not only for clinical intervention in patients with leukemia but also as chemical tools to further dissect BCR-Abl kinase-linked signaling pathways in tissue culture cells (11). However, efforts to analyze the effects of monospecific inhibition of BCR-Abl kinase have been complicated by cross-reactivity of ATP-competitive Abl inhibitors with other kinases. For example, in addition to inhibiting c-Abl and BCR-Abl, STI-571 and nilotinib also potently inhibit c-Kit, platelet-derived growth factor receptor, and DDR1, whereas dasatinib potently inhibits all of these kinases as well as the Src family, Tec family, and KDR kinases (12). The multitargeted nature of these ATP-competitive inhibitors makes it difficult to assign a particular biological effect to inhibition of a specific kinase target.We previously reported the discovery of the first non-ATP site-monoselective BCR-Abl inhibitor (GNF-2), which targets not only wild type BCR-Abl but also many clinically relevant STI-571-resistant mutants either alone or in combination with other BCR-Abl inhibitors (13). Molecular modeling, site-directed mutagenesis, competition assays, NMR spectroscopy, and protein crystallography were used to determine that GNF-2 binds to a myristate-binding site in the C-lobe of the c-Abl kinase domain (Fig. 1, B and C) (3). The discovery of GNF-2 was the first demonstration that c-Abl kinase activity could be pharmacologically modulated by an inhibitor that binds outside the ATP or substrate binding sites. Although it remained unclear how GNF-2 is capable of inhibiting c-Abl upon binding to the myristate-binding site, we speculated that GNF-2 probably mimics the function of the N-myristoyl residue in c-Abl. Here, we investigated the effects of GNF-2 on Abl family members with the goals of providing further insights into the mechanism of GNF-2 function and laying the foundation to utilize GNF-2 as a tool to investigate c-Abl- and Arg-linked cellular processes. 相似文献
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Ellen Weisberg Chengcheng Meng Abigail E. Case Hong L. Tiv Prafulla C. Gokhale Anthia A. Toure Sara Buhrlage Xiaoxi Liu Jinhua Wang Nathanael Gray Richard Stone Sophia Adamia Eric Winer Martin Sattler James D. Griffin 《Journal of cellular and molecular medicine》2020,24(3):2145-2156
Mutations in the E3 ubiquitin ligase CBL, found in several myeloid neoplasms, lead to decreased ubiquitin ligase activity. In murine systems, these mutations are associated with cytokine‐independent proliferation, thought to result from the activation of hematopoietic growth receptors, including FLT3 and KIT. Using cell lines and primary patient cells, we compared the activity of a panel of FLT3 inhibitors currently being used or tested in AML patients and also evaluated the effects of inhibition of the non‐receptor tyrosine kinase, SYK. We show that FLT3 inhibitors ranging from promiscuous to highly targeted are potent inhibitors of growth of leukaemia cells expressing mutant CBL in vitro, and we demonstrate in vivo efficacy of midostaurin using mouse models of mutant CBL. Potentiation of effects of targeted FLT3 inhibition by SYK inhibition has been demonstrated in models of mutant FLT3‐positive AML and AML characterized by hyperactivated SYK. Here, we show that targeted SYK inhibition similarly enhances the effects of midostaurin and other FLT3 inhibitors against mutant CBL‐positive leukaemia. Taken together, our results support the notion that mutant CBL‐expressing myeloid leukaemias are highly sensitive to available FLT3 inhibitors and that this effect can be significantly augmented by optimum inhibition of SYK kinase. 相似文献