排序方式: 共有5条查询结果,搜索用时 0 毫秒
1
1.
Crisona NANCY J. Allen KAREN D. Strohman RICHARD C. 《Journal of muscle research and cell motility》1998,19(1):43-51
Skeletal muscle has the remarkable capacity to regenerate new muscle fibres in the event of injury or disease. This capacity lies in the satellite cells, which are myogenic stem cells residing in adult muscle. While the signals that activate satellite cells to divide in vivo are not fully understood, satellite cells grown in culture respond to the mitogenic action of fibroblast growth factor (FGF). Satellite cells from the dystrophic mdx mouse are more sensitive to FGF in culture than satellite cells from normal mice. In this study we investigated the basis for this heightened sensitivity of mdx satellite cells to FGF by measuring the number and affinity of protein and heparan sulphate proteoglycan (HSPG) receptors for FGF. We found that HSPG receptors were elevated over four-fold in the mdx cells compared with cells from normal animals. We supported this observation by measuring the synthesis of heparan sulphate (HS) and chondroitin sulphate (CS) by satellite cells in culture. Mdx satellite cells synthesized approximately ten times more of these sulphated glycosaminoglycans (GAGs) than did normal cells. For muscle fibroblasts, however, we found no significant difference in the number or affinity of protein or HSPG receptors, or in the amount of sulphated GAGs synthesized, between normal and mdx cells. We propose that the increase in FGF HSPG receptors is the basis for the heightened response of mdx satellite cells to FGF in culture and may reflect exposure of the cells to growth factors in the degenerating mdx muscleThis revised version was published online in July 2005 with corrections to the Cover Date. 相似文献
2.
Chirality sensing by Escherichia coli topoisomerase IV and the mechanism of type II topoisomerases 总被引:3,自引:0,他引:3 下载免费PDF全文
Stone MD Bryant Z Crisona NJ Smith SB Vologodskii A Bustamante C Cozzarelli NR 《Proceedings of the National Academy of Sciences of the United States of America》2003,100(15):8654-8659
Escherichia coli topoisomerase (Topo) IV is an essential type II Topo that removes DNA entanglements created during DNA replication. Topo IV relaxes (+) supercoils much faster than (-) supercoils, promoting replication while sparing the essential (-) supercoils. Here, we investigate the mechanism underlying this chiral preference. Using DNA binding assays and a single-molecule DNA braiding system, we show that Topo IV recognizes the chiral crossings imposed by the left-handed superhelix of a (+) supercoiled DNA, rather than global topology, twist deformation, or local writhe. Monte Carlo simulations of braid, supercoil, and catenane configurations demonstrate how a preference for a single-crossing geometry during strand passage can allow Topo IV to perform its physiological functions. Single-enzyme braid relaxation experiments also provide a direct measure of the processivity of the enzyme and offer insight into its mechanochemical cycle. 相似文献
3.
Lisa Postow Nancy J. Crisona Brian J. Peter Christine D. Hardy Nicholas R. Cozzarelli 《Proceedings of the National Academy of Sciences of the United States of America》2001,98(15):8219-8226
The unwinding of the parental DNA duplex during replication causes a positive linking number difference, or superhelical strain, to build up around the elongating replication fork. The branching at the fork and this strain bring about different conformations from that of (-) supercoiled DNA that is not being replicated. The replicating DNA can form (+) precatenanes, in which the daughter DNAs are intertwined, and (+) supercoils. Topoisomerases have the essential role of relieving the superhelical strain by removing these structures. Stalled replication forks of molecules with a (+) superhelical strain have the additional option of regressing, forming a four-way junction at the replication fork. This four-way junction can be acted on by recombination enzymes to restart replication. Replication and chromosome folding are made easier by topological domain barriers, which sequester the substrates for topoisomerases into defined and concentrated regions. Domain barriers also allow replicated DNA to be (-) supercoiled. We discuss the importance of replicating DNA conformations and the roles of topoisomerases, focusing on recent work from our laboratory. 相似文献
4.
Preferential relaxation of positively supercoiled DNA by E. coli topoisomerase IV in single-molecule and ensemble measurements 总被引:9,自引:0,他引:9 下载免费PDF全文
Crisona NJ Strick TR Bensimon D Croquette V Cozzarelli NR 《Genes & development》2000,14(22):2881-2892
We show that positively supercoiled [(+) SC] DNA is the preferred substrate for Escherichia coli topoisomerase IV (topo IV). We measured topo IV relaxation of (-) and (+) supercoils in real time on single, tethered DNA molecules to complement ensemble experiments. We find that the preference for (+) SC DNA is complete at low enzyme concentration. Otherwise, topo IV relaxed (+) supercoils at a 20-fold faster rate than (-) supercoils, due primarily to about a 10-fold increase in processivity with (+) SC DNA. The preferential cleavage of (+) SC DNA in a competition experiment showed that substrate discrimination can take place prior to strand passage in the presence or absence of ATP. We propose that topo IV discriminates between (-) and (+) supercoiled DNA by recognition of the geometry of (+) SC DNA. Our results explain how topo IV can rapidly remove (+) supercoils to support DNA replication without relaxing the essential (-) supercoils of the chromosome. They also show that the rate of supercoil relaxation by topo IV is several orders of magnitude faster than hitherto appreciated, so that a single enzyme may suffice at each replication fork. 相似文献
5.
Dekker NH Rybenkov VV Duguet M Crisona NJ Cozzarelli NR Bensimon D Croquette V 《Proceedings of the National Academy of Sciences of the United States of America》2002,99(19):12126-12131
The topology of cellular DNA is carefully controlled by enzymes called topoisomerases. By using single-molecule techniques, we monitored the activity of two type IA topoisomerases in real time under conditions in which single relaxation events were detected. The strict one-at-a-time removal of supercoils we observed establishes that these enzymes use an enzyme-bridged strand-passage mechanism that is well suited to their physiological roles and demonstrates a mechanistic unity with type II topoisomerases. 相似文献
1