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41.
An autonomously replicating sequence of pSRI plasmid is effective in two yeast species, Zygosaccharomyces rouxii and Saccharomyces cerevisiae 总被引:2,自引:0,他引:2
The autonomously replicating sequences (ARSs) of pSR1, a cryptic circular DNA plasmid detected in a strain of Zygosaccharomyces rouxii, were delimited by subcloning and deletion analysis and by the isolation of nucleotide substitution mutations. A 30 base-pair (bp) sequence from inverted repeat 1 (IR1) and presumably the same region from IR2 of pSR1 functions as an ARS in the native host, Z. rouxii, and in a heterologous host, Saccharomyces cerevisiae. Thus, pSR1 has two ARSs per molecule, either of which is sufficient for replication of the plasmid molecule in both hosts. These hosts, however, respond differently to nucleotide substitutions in the 30 bp sequence, suggesting that the sequences required for ARS function in the two organisms are not exactly the same. In addition, a 137 bp sequence that overlaps the 30 bp sequence by 11 bp also functions as an ARS in Z. rouxii but not in S. cerevisiae. However, this 137 bp sequence enhances the stability of plasmids carrying the pSR1 ARS in S. cerevisiae. The 30 bp and 137 bp sequences each contain a single copy of the 11 bp ARS consensus sequence, which is essential for ARS function in S. cerevisiae. Small insertions between the 11 bp overlapping region and the 11 bp ARS consensus sequence showed that a proper distance between these two 11 bp sequences is essential for the ARS function of the 30 bp sequence. Point mutations that inactivate ARS function show that the ARS consensus sequence, as well as a short A:T segment in the overlapping sequence, is required for the ARS function of the 30 bp sequence. 相似文献
42.
Ultrastructure of neurofibrillary tangles in Alzheimer's disease 总被引:3,自引:0,他引:3
T Miyakawa S Katsuragi K Araki T Hashimura T Kimura R Kuramoto 《Virchows Archiv. B, Cell pathology including molecular pathology》1989,57(4):267-273
The ultrastructure of neurofibrillary tangles (NFT) was examined by electron microscopy. The fibrils of NFT seemed to consists of about eight protofilaments consisting of globular subunits; these protofilaments were helically wound in a longitudinal direction. The fibrils of NFT had hollow structures at their centers surrounded by the eight globular subunits. The subunits were tightly connected in the narrow parts of the fibril, but more loosely connected in the wider parts. From these findings, it seemed that the fibrils of NFT consist of a twisted tubule having periodical constrictions and is made up of eight helically wound protofilaments, forming globular subunits. 相似文献
43.
A Kikuchi T Sasaki S Araki Y Hata Y Takai 《The Journal of biological chemistry》1989,264(16):9133-9136
The smg-21 GTP-binding protein (smg p21) has the same effector domain as the ras proteins (ras p21s) and is identical with the proteins of the rap1A and Krev-1 genes. In this paper, two proteins stimulating the GTPase activity of smg p21 are partially purified from bovine brain cytosol. These proteins, designated as smg p21 GTPase-activating protein (GAP) 1 and 2, are separated from a c-ras p21 GAP described previously by column chromatographies. smg p21 GAP1 and -2 stimulate the GTPase activity of only smg p21 but not that of c-Ha-ras p21 or the rho and smg-25A GTP-binding proteins. smg p21 GAP1 or -2 does not stimulate the dissociation of guanosine 5'-3-O-(thio)triphosphate or GDP from smg p21. smg p21 GAP1 or -2 themselves do not have GTP/GDP binding or GTPase activity. The Mr values of smg p21 GAP1 and -2 are estimated to be 250-400 x 10(3) and 80-100 x 10(3) by gel filtration and sucrose density gradient ultracentrifugation, respectively. The activity of smg p21 GAP1 and -2 is killed by tryptic digestion or heat boiling. These results indicate that bovine brain contains two smg p21 GAPs in addition to c-ras p21 GAP. 相似文献
44.
45.
Severe combined immune deficiency (scid) mice are assumed to have two types of abnormalities: one is high radiosensitivity and the other is abnormal recombination
in immunoglobulin and T-cell receptor genes. The human chromosome 8 q1.1 region has an ability to complement the scid aberrations. Moreover, the localization of the subunit DNA-dependent protein kinase [DNA-PKcs] participating in DNA double-strand break repair in the same locus was clarified. In scid mouse cells, the number of DNA-PKcs products and extent of DNA-PK activity remarkably decrease. These observations gave rise to the assumption that DNA-PKcs is the scid factor itself. In order to determine whether the DNA-PK
cs
gene is the scid gene, we isolated the mouse DNA-PK
cs
gene and investigated its chromosomal locus by fluorescence in situ hybridization (FISH). Consequently, it became clear that
the mouse DNA-PK
cs
gene existed in the centromeric region of mouse chromosome 16, determined by cross-genetic study, as a scid locus. This finding strongly suggests that mouse DNA-PK
cs
is the scid gene.
Received: 22 March 1996 相似文献
46.
We found an intra-acrosomal antigen of about 155,000 daltons (155 kDa) in a survey using the monoclonal antibody MC101 raised against mouse cauda epididymal spermatozoa. Morphological studies by means of indirect immunofluorescence and immunogold electron microscopy localized the antigen to the cortex region of the anterior acrosome. Avidin biotin complex immunocytochemistry initially demonstrated a faint signal at the anterior acrosome in the testis spermatozoa that increased in intensity as the sperm moved toward the distal epididymis. This incremental immunoreactivity was also confirmed by immunoblotting following one-dimensional SDS-PAGE. The 155 kDa protein band was immunostained, and it was much more intense in the cauda epididymal than in the caput and corpus epididymal spermatozoa. Only a trace or no immunostain was evident in the caput or testis spermatozoa. The antigen localization did not change during passage through the epididymis, being confined at the cortex region of the anterior acrosome. The epididymal epithelial cells were not immunostained. These findings suggested that the 155 kDa protein is biochemically modified, further implying that the biochemical alteration of intra-acrosomal material is involved in sperm maturation in the epididymis. © 1995 wiley-Liss, Inc. 相似文献
47.
rac p21 is involved in insulin-induced membrane ruffling and rho p21 is involved in hepatocyte growth factor- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced membrane ruffling in KB cells. 总被引:17,自引:4,他引:13 下载免费PDF全文
T Nishiyama T Sasaki K Takaishi M Kato H Yaku K Araki Y Matsuura Y Takai 《Molecular and cellular biology》1994,14(4):2447-2456
Insulin and hepatocyte growth factor (HGF) induced morphologically different membrane rufflings in KB cells. Insulin-induced membrane ruffling was inhibited by microinjection of rho GDI, an inhibitory GDP/GTP exchange regulator for both rho p21 and rac p21 small GTP-binding proteins, but not inhibited by microinjection of botulinum exoenzyme C3, known to selectively ADP-ribosylate rho p21 and to impair its function. This rho GDI action was prevented by comicroinjection with guanosine 5'-(3-O-thio)triphosphate (GTP gamma S)-bound rac1 p21. In contrast, HGF-induced membrane ruffling was inhibited by microinjection of rho GDI or C3. This rho GDI action was prevented by comicroinjection with GTP gamma S-bound rhoA p21, and this C3 action was prevented by comicroinjection with GTP gamma S-bound rhoAIle-41 p21, which is resistant to C3. Microinjection of either GTP gamma S-bound rac1 p21 or rhoA p21 alone induced membrane ruffling in the absence of the growth factors. The rac1 p21-induced membrane ruffling was morphologically similar to the insulin-induced kind, whereas rhoA p21-induced ruffling was apparently different from both the insulin- and HGF-induced kinds. Membrane ruffling was also induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C-activating phorbol ester, but not by Ca2+ ionophore or microinjection of a dominant active Ki-ras p21 mutant (Ki-rasVal-12 p21). The phorbol ester-induced membrane ruffling was morphologically similar to the rhoA p21-induced kind and inhibited by microinjection of rho GDI or C3. These results indicate that rac p21 and rho GDI are involved in insulin-induced membrane ruffling and that rho p21 and rho GDI are involved in HGF- and phorbol ester-induced membrane rufflings. 相似文献
48.
K. Funase K. Imanaka Y. Nishihira H. Araki 《European journal of applied physiology and occupational physiology》1994,69(1):21-25
In the present study, we investigated whether weak (10% of maximal voluntary contraction) tonic dorsiflexion (DF) and plantarflexion (PF) affects the two conventional parameters used for evaluating the excitability of the soleus motoneuron (MN) pool, i.e. the ratio of the threshold of H-reflex to that of M-response (Hth:Mth) and the ratio of the maximal amplitude of H-reflex to that of M-response (Hmax:Mmax) in human subjects. The results showed that the Hmax:Mmax decreased during DF and increased during PF compared with that during rest, whereas no clear alteration was observed in Hth:Mth. These results are consistent with the scheme proposed by earlier workers, who have argued that neither inhibitory nor facilitatory effects of the conditioning stimulus apply to specific spinal reflex circuits occurring around the threshold of the test H-reflex. It is suggested, therefore, that the conventional use of the Hth:Mth ratio as a parameter reflecting the excitability of the MN pool should be reconsidered. 相似文献
49.
Soh Chang-Ho; Kamiya Yuji; Yoshida Shigeo; Yamane Hisakazu; Takahashi Nobutaka 《Plant & cell physiology》1994,35(7):1037-1042
Oryzains, cysteine proteinases of rice seeds, are induced byGA3 in germinating rice seeds [Abe et al. (1987) Agric. Biol.Chem. 51: 1509]. The effects of GA1, GA3, GA4, GA9, and GA20on the production of oryzain and -amylase were investigatedin embryoless half- and whole-seeds of rice (cv. Nipponbare).When gibberellins (GAs) were incubated with embryoless half-seeds,GA1, GA3 and GA4 induced oryzain and -amylase, but GA9, andGA20 did not. GA9 and GAM induced oryzain and -amylase productionin whole seeds, but this production was inhibited by the simultaneousapplication of prohexadione, an inhibitor of 2ß- and3ß-hydroxylation of GAs. Prohexadione did not inhibitthe activities of oryzain and -amylase induced by GA1. Theseresults suggest that GAs possessing the 3ß-hydroxylgroup induce activities of oryzain and -amylase in rice seedsand that GA9 and GA20 have activity only after they are convertedmetabolically to active GAs, probably GA4 and GA1, respectively.GA1, was more active than GA4 in both half seeds and wholeseeds incubation. Oryzain and -amylase activities induced byGA4 were significantly inhibited in the presence of 104M prohexadione. This suggests that the conversion of GA1, toGA4 (13-hydroxylation) might be inhibited at a high dose ofprohexadione in whole seeds.
4Present address: Institute of Food Development, Kyung Hee University,Suwon 449-701, Korea 相似文献
50.
Shukuro Araki Shigehiro Yi Tatsufumi Murakami Susumu Watanabe Shinichi Ikegawa Kiyoshi Takahashi Ken-ichi Yamarnura 《Molecular neurobiology》1994,8(1):15-23
To analyze the pathologic processes of amyloid deposition in type I familial amyloidotic polyneuropathy (FAP), mice were made
transgenic by introducing the human mutant transthyretin (TTR) gene(MT-hMet 30). An inbred strain of mouse, C57 BL/6, was
chosen. Transgenic mice were killed using ether anesthesia at 3-mo intervals up to 24 mo after birth. In these transgenic
mice, amyloid deposition started in the gastrointestinal tract, cardiovascular system, and kidneys and extended to various
other organs and tissues with advancing age. The pattern of amyloid deposition was similar to that observed in human autopsy
cases of FAP, except for its absence in the choroid plexus and in the peripheral and autonomic nervous systems.
We extracted the amyloid fibrils from kidneys of these mice with a human mutant TTR gene and analyzed them immunochemically
and electronmicroscopically. Deposited amyloid was shown to be composed of human mutant TTR and mouse serum amyloid P component.
Amyloid fibril from transgenic mice was morphologically and immunohistochemically similar to that of human FAP.
The most striking pathologic feature of the transgenic mice was the absence of amyloid deposition in the peripheral and autonomic
nervous tissues. Thus, other intrinsic factors may be involved in amyloid deposition in the nervous tissues of human FAP. 相似文献