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81.
82.
Inhibition of endoplasmic reticulum (ER)-to-Golgi transport induces relocalization of binding protein (BiP) within the ER to form the BiP bodies. 总被引:11,自引:6,他引:5
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Immunofluorescence staining of yeast cells with anti-binding protein (BiP) antibodies shows uniform staining of the endoplasmic reticulum (ER). We have found that overproduction of Sec12p, an ER membrane protein, causes a change of BiP distribution within the cell. Upon induction of Sec12p by the GAL1 promoter, the staining pattern of BiP turns into bright dots scattering in the cell, whereas the staining of Sec12p remains to be the typical ER figure. Overproduction of other ER membrane proteins, HMG-CoA reductase or Sed4 protein, does not induce such relocalization of BiP. Pulse-chase experiments and electron microscopy have revealed that the overproduction of Sec12p inhibits protein transport from the ER to the Golgi apparatus. When the transport is arrested by one of the sec mutations that block the ER-to-Golgi step at the restrictive temperature, the BiP staining also changes into the punctate pattern. In contrast, the sec mutants that block later or earlier steps of the secretory pathway do not induce such change of BiP localization. These observations indicate that relocalization of BiP is caused by the inhibition of ER-to-Golgi transport. Using immunoelectron microscopy, we have found that the punctate staining is because of the accumulation of BiP in the restricted region of the ER, which we propose to call the "BiP body." This implicates existence of ER subdomains in yeast. A vacuolar protein, proteinase A, appears to colocalize in the BiP body when the ER-to-Golgi transport is blocked, suggesting that the BiP body may have a role as the site of accumulation of cargo molecules before exit from the ER. 相似文献
83.
The behavior of cell nuclei, mitochondrial nucleoids (mt-nucleoids) and plastid nucleoids (ptnucleoids) was studied in the
root apical meristem of Arabidopsis thaliana. Samples were embedded in Technovit 7100 resin, cut into thin sections and stained with 4′-6-diamidino-2-phenylindole for
light-microscopic autoradiography and microphotometry. Synthesis of cell nuclear DNA and cell division were both active in
the root apical meristem between 0 μm and 300 μm from the central cells. It is estimated that the cells generated in the lower
part of the root apical meristem enter the elongation zone after at least four divisions. Throughout the entire meristematic
zone, individual cells had mitochondria which contained 1–5 mt-nucleoids. The number of mitochondria increased gradually from
65 to 200 in the meristem of the central cylinder. Therefore, throughout the meristem, individual mitochondria divided either
once or twice per mitotic cycle. By contrast, based on the incorporation of [3H]thymidine into organelle nucleoids, syntheses of mitochondrial DNA (mtDNA) and plastid DNA (ptDNA) occurred independently
of the mitotic cycle and mainly in a restricted region (i.e., the lower part of the root apical meristem). Fluorimetry, using
a videointensified microscope photon-counting system, revealed that the amount of mtDNA per mt-nucleoid in the cells in the
lower part of the meristem, where mtDNA synthesis was active, corresponded to more than 1 Mbp. By contrast, in the meristematic
cells just below the elongation zone of the root tip, the amount of mtDNA per mt-nucleoid fell to approximately 170 kbp. These
findings strongly indicate that the amount of mtDNA per mitochondrion, which has been synthesized in the lower part of the
meristem, is gradually reduced as a result of continual mitochondrial divisions during low levels of mtDNA synthesis. This
phenomenon would explain why differentiated cells in the elongation zone have mitochondria that contain only extremely small
amounts of mtDNA.
This work was supported by a Grant-in Aid (T.K.) for Special Research on Priority Areas (Project No. 02242102, Cellular and
Molecular Basis for Reproduction Processes in Plants) from the Ministry of Education, Science and Culture of Japan and by
a Grant-in Aid (T.K.) for Original and Creative Research Project on Biotechnology from the Research Council, Ministry of Agriculture,
Forestry and Fisheries of Japan. 相似文献
84.
Heat production, oxygen consumption, and lipolysis in isolated interscapular brown adipocytes from the rat were investigated. Epinephrine, norepinephrine, and isoproterenol increased heat production in a concentration-dependent manner, showing, about 6-, 4-, and 5-fold higher effects than controls, respectively. The concentration of isoproterenol for threshold heat production and glycerol release were 10(-10) M and 10(-9) M, respectively. The fact that 10(-9) M isoproterenol increased heat production by about 3-fold while glycerol release had no effect at all indicates that calorimetry is more appropriate for investigation of brown adipocytes. At least the method is more sensitive than that of measuring glycerol release. 相似文献
85.
Acetyl esterase production was detected in a wood-rotting fungus,Coriolus versicolor, by the formation of a clear zone on a double layer agar plate containing glucose β-d-pentaacetate. Two polysaccharide acetates, carboxymethyl cellulose acetate and xylan acetate, also served as detectable substrates
in place of glucose acetate to form clear zone. In an esterase assay, this fungal esterase showed a higher specificity to
acetylxylan than did porcine liver esterase, indicating that it is an acetylxylan esterase. 相似文献
86.
A Novel Class of Herbicides (Specific Inhibitors of Imidazoleglycerol Phosphate Dehydratase) 总被引:4,自引:2,他引:2
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Mori I Fonne-Pfister R Matsunaga S Tada S Kimura Y Iwasaki G Mano J Hatano M Nakano T Koizumi S Scheidegger A Hayakawa K Ohta D 《Plant physiology》1995,107(3):719-723
A new mode of herbicidal action was established by finding specific inhibitors of imidazoleglycerol phosphate dehydratase, an enzyme of histidine (His) biosynthesis. Three triazole phosphonates inhibited the reaction of the enzyme with Ki values of 40 [plus or minus] 6.5, 10 [plus or minus] 1.6, and 8.5 [plus or minus] 1.4 nM, respectively, and were highly cytotoxic to cultured plant cells. This effect was completely reversed by the addition of His, proving that the cytotoxicity was primarily caused by the inhibition of His biosynthesis. These inhibitors showed wide-spectrum, postemergent herbicidal activity at application rates ranging from 0.05 to 2 kg/ha. 相似文献
87.
88.
Masaru Nakano Keizo Hosokawa Tomo Oomiya Saburo Yamamura 《Plant Cell, Tissue and Organ Culture》1995,41(3):221-227
A protoplast-to-plant system was developed in Gentiana using a gellan gum-embedding culture. Viable protoplasts could be routinely isolated from in vitro-grown plantlets, and they were embedded in 0.2% gellan gum-solidified B5 medium containing 2 mg l-1 NAA, 0.1 mg l-1 TDZ, 0.1 M sucrose and 0.4 M mannitol. Weekly addition of fresh liquid medium was essential for preventing cell browning. Colony growth was promoted by lowering mannitol concentration of the culture media after one month, and visible colonies were produced after 2 months of culture. Shoot regeneration from protoplast-derived calluses was stimulated by 1 to 10 mg l-1 TDZ in combination with 0.1 mg l-1 NAA. Protoplast-derived plants were recovered following rooting of the shoots in plant growth regulator-free medium and they were successfully transferred to soil.Abbreviations BA
benzylaminopurine
- FDA
fluorescein diacetate
- FW
fresh weight
- MES
2-N-morpholinoethane sulfonic acid
- NAA
-naphthaleneacetic acid
- TDZ
N-1,2,3-thiadiazol-5-yl-N-phenylurea (also called thidiazuron) 相似文献
89.
90.