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31.
32.
Tissue type-specific expression of intermediate filament proteins in a cultured epithelial cell line from bovine mammary gland 总被引:15,自引:11,他引:15 下载免费PDF全文
Different clonal cell lines have been isolated from cultures of mammary gland epithelium of lactating cow’s udder and have been grown in culture media containing high concentrations of hydrocortisone, insulin, and prolactin. These cell (BMGE+H), which grow in monolayers of typical epithelial appearance, are not tightly packed, but leave intercellular spaces spanned by desmosomal bridges. The cells contain extended arrays of cytokeratin fibrils, arranged in bundles attached to desmosomes. Gel electophoresis show that they synthesize cytokeratins similar, if not identical, to those found in bovine epidermis and udder, including two large (mol wt 58,500 and 59,000) and basic (pH range: 7-8) and two small (mol wt 45,500 and 50,000) and acidic (pH 5.32 and 5.36) components that also occur in phosphorylated forms. Two further cytokeratins of mol wts 44,000 (approximately pH 5.7) and 53,000 (pH 6.3) are detected as minor cytokeratins in some cell clones. BMGE+H cells do not produce vimentin filaments as determined by immunofluorescence microscopy and gel electrophoresis. By contrast, BMGE-H cells, which have emerged from the same original culture but have been grown without hormones added, are not only morphologically different, but also contain vimentin filaments and a different set of cytokeratins, the most striking difference being the absence of the two acidic cytokeratins of mol wt 50,000 and 45,500. Cells of the BMGE+H line are characterized by an unusual epithelial morphology and represent the first example of a nonmalignant permanent cell line in vitro that produces cytokeratin but not vimentin filaments. The results show that (a) tissue-specific patterns of intermediate filament expression can be maintained in permanent epithelial cell lines in culture, at least under certain growth conditions; (b) loss of expression of relatively large, basic cytokeratins is not an inevitable consequence of growth of epithelial cells in vitro. Our results further show that, during culturing, different cell clones with different cytoskeletal composition can emerge from the same cell population and suggest that the presence of certain hormones may have an influence on the expression of intermediate filament proteins. 相似文献
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35.
EDC3 phosphorylation regulates growth and invasion through controlling P‐body formation and dynamics
Jeremiah J Bearss Sathish KR Padi Neha Singh Marina CardoVila Jin H Song Ghassan Mouneimne Nikita Fernandes Yang Li Matthew R Harter Jaime MC Gard Anne E Cress Wolfgang Peti Andrew DL Nelson J Ross Buchan Andrew S Kraft Koichi Okumura 《EMBO reports》2021,22(4)
Regulation of mRNA stability and translation plays a critical role in determining protein abundance within cells. Processing bodies (P‐bodies) are critical regulators of these processes. Here, we report that the Pim1 and 3 protein kinases bind to the P‐body protein enhancer of mRNA decapping 3 (EDC3) and phosphorylate EDC3 on serine (S)161, thereby modifying P‐body assembly. EDC3 phosphorylation is highly elevated in many tumor types, is reduced upon treatment of cells with kinase inhibitors, and blocks the localization of EDC3 to P‐bodies. Prostate cancer cells harboring an EDC3 S161A mutation show markedly decreased growth, migration, and invasion in tissue culture and in xenograft models. Consistent with these phenotypic changes, the expression of integrin β1 and α6 mRNA and protein is reduced in these mutated cells. These results demonstrate that EDC3 phosphorylation regulates multiple cancer‐relevant functions and suggest that modulation of P‐body activity may represent a new paradigm for cancer treatment. 相似文献
36.
TL Kieft EM Murphy DL Haldeman PS Amy BN Bjornstad EV McDonald DB Ringelberg DC White J Stair RP Griffiths TC Gsell WE Holben DR Boone 《Microbial ecology》1998,36(3):336-348
Abstract Two chronosequences of unsaturated, buried loess sediments, ranging in age from <10,000 years to >1 million years, were investigated to reconstruct patterns of microbial ecological succession that have occurred since sediment burial. The relative importance of microbial transport and survival to succession was inferred from sediment ages, porewater ages, patterns of abundance (measured by direct counts, counts of culturable cells, and total phospholipid fatty acids), activities (measured by radiotracer and enzyme assays), and community composition (measured by phospholipid fatty acid patterns and Biolog substrate usage). Core samples were collected at two sites 40 km apart in the Palouse region of eastern Washington State, near the towns of Washtucna and Winona. The Washtucna site was flooded multiple times during the Pleistocene by glacial outburst floods; the Winona site elevation is above flood stage. Sediments at the Washtucna site were collected from near surface to 14.9 m depth, where the sediment age was approximately 250 ka and the porewater age was 3700 years; sample intervals at the Winona site ranged from near surface to 38 m (sediment age: approximately 1 Ma; porewater age: 1200 years). Microbial abundance and activities declined with depth at both sites; however, even the deepest, oldest sediments showed evidence of viable microorganisms. Same-age sediments had equal quantities of microorganisms, but different community types. Differences in community makeup between the two sites can be attributed to differences in groundwater recharge and paleoflooding. Estimates of the microbial community age can be constrained by porewater and sediment ages. In the shallower sediments (<9 m at Washtucna, <12 m at Winona), the microbial communities are likely similar in age to the groundwater; thus, microbial succession has been influenced by recent transport of microorganisms from the surface. In the deeper sediments, the populations may be considerably older than the porewater ages, since microbial transport is severely restricted in unsaturated sediments. This is particularly true at the Winona site, which was never flooded. 相似文献
37.
Selective neutrality of glucose-6-phosphate dehydrogenase allozymes in Escherichia coli 总被引:2,自引:0,他引:2
Six naturally occurring alleles representing four electromorphs of the
enzyme glucose-6-phosphate dehydrogenase were transferred by P1- mediated
transduction from natural isolates of Escherichia coli into the genetic
background of E. coli K12 and were studied in pairwise competition in
chemostats limited for glucose in order to estimate differences in growth
rate associated with the alleles. Although the level of resolution of such
experiments is a growth rate differential of approximately 0.002 h-1, no
significant differences among the strains were found. Studies of apparent
Km and Vmax in crude enzyme extracts of the strains also failed to reveal
any significant differences among the electromorphs. These results support
the view that the alleles are selectively neutral or nearly neutral under
these conditions.
相似文献
38.
Chromosomal DNA from 23 closely related, pathogenic strains of Escherichia
coli was digested and probed for the insertion sequences IS1, IS2, IS4,
IS5, and IS30. Under the assumption that elements residing in DNA
restriction fragments of the same apparent length are identical by descent,
parsimony analysis of these characters yielded a unique phylogenetic tree.
This analysis not only distinguished among bacterial strains that were
otherwise identical in their biochemical characteristics and enzyme
electrophoretic mobilities, but certain aspects of the topology of the tree
were consistent across several unrelated insertion elements. The
distribution of IS elements was then reexamined in light of the inferred
phylogenetic relationships to investigate the biological properties of the
elements, such as rates of insertion and deletion, and to discover apparent
recombinational events. The analysis shows that the pattern of distribution
of insertion elements in the bacterial genome is sufficiently stable for
epidemiological studies. Although the rate of recombination by conjugation
has been postulated to be low, at least two such events appear to have
taken place.
相似文献
39.
Sequence, organization, and evolution of the A+T region of Drosophila melanogaster mitochondrial DNA 总被引:2,自引:0,他引:2
The long (4.6-kb) A+T region of Drosophila melanogaster mitochondrial DNA
has been cloned and sequenced. The A+T region is organized in two large
arrays of tandemly repeated DNA sequence elements, with nonrepetitive
intervening and flanking sequences comprising only 22% of its length. The
first repeat array consists of five repeats of 338-373 bp. The second
consists of four intact 464-bp repeats and a fifth partial repeat of 137
bp. Three DNA sequence elements are found to be highly conserved in D.
melanogaster and in several Drosophila species with short A+T regions.
These include a 300-bp DNA sequence element that overlaps the DNA
replication origin and two thymidylate stretches identified on opposite DNA
strands. We conclude that the length heterogeneity observed in the A+T
regulatory region in mitochondrial DNAs from the genus Drosophila results
from the expansion (and contraction) of the number of repeated DNA sequence
elements. We also propose that the 300-bp conserved DNA sequence element,
in conjunction with another primary sequence determinant, perhaps the
adjacent thymidylate stretch, functions in the regulation of mitochondrial
DNA replication.
相似文献
40.