• Korean Journal of Microbiology
• /
• v.38 no.4
• /
• pp.218-218
• /
• 2002

Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

• Applied Microscopy
• /
• v.27 no.2
• /
• pp.131-144
• /
• 1997

The present study was carried out to investigate the processes of the ultrastructural differentiation and the acetylcholinesterase (AChE) activities of the developing rat retina. The results are as follows. The retina of fetal rat on the 13th day of gestation showed the early stage of differentiation. Briefly, there appeared dividing chromosomes, the plentiful free ribosomes, and the high ratio of nucleus to cytoplasm. The reaction products by AChE were localized at the membrane of endoplasmic reticulum and on the outer membrane of nucleus. Ultrastructures and AChE activities in the retina of the fetal rats on the 18th day of gestation were similar to those of the prior stages, except the appearence of rough endoplasmic reticulum and Golgi apparatus. According to the ultrastructural observations, the rat retina was still in immature state at birth, but the pigment epithelial cells were fully differentiated, e. g. the increase of melanin granules, the development of mitochondria and Golgi apparatus. The AChE activity was weekly detected. The differentiated retinal layers and the outer segment of photoreceptor cells were observed on the 7th postnatal day. And the pigment epithelium appeared to be fully differentiated. On the 14th postnatal day, rat retina were completely differentiated. In other words, the rat retina was characterized by the prominent outer segments, phagocytosed residues in the pigment epithelium, and the localization of reaction products by AChE in the synapses. In conclusion, the differentiation of rat retina is charaterized by the changes of cell shape, the increase of retinal layers, and the alterations of AChE activities. It seems that rat retina is to be functional from 2 weeks of birth onward, coinciding with the eye opening of the juvenile rats.

• Applied Microscopy
• /
• v.13 no.1
• /
• pp.71-84
• /
• 1983

This study was made to investigate the ultrastructural changes of the hepatocyte of the maternal liver, and fetal liver by Actinomycin D in Wistar rats at the stage of pregnancy. Peritoneal injection of Actinomycin D to rats carried out gestation day 7 to 9 at the level of $15{\mu}g(11.5{\mu}g/100g$ body wt.), $20{\mu}g(15.8{\mu}g/100g$ body wt.) on each day. Treated animals with saline only were used for controls. Animals were sacrificed on day 15 of gestation. On electron microscopic examination, the hepatocytes of maternal liver given Actinomycin D $15{\mu}g$/ml had evidence of serious cellular damage, for example, hypertrophy of rough endoplasmic reticulum, loss in nucleolar osmiophilia, swelling of Golgi apparatus and change of mitochondrial structure. Maternal liver given Actinomycin D $20{\mu}g/ml$ shown similar changes to that of the $15{\mu}g/ml$ treated animals. But mitochondria of this group were not changed than that of $15{\mu}g/ml$ treated group. In the hepatocytes of fetal liver, changes were more pronounced. The drug produced alteration in nuclei and cytoplasm. The rough endoplasmic reticulum was swollen and there were ribosomes detachement. In addition, damages of mitochondria, Golgi apparatus were detected.

• Applied Microscopy
• /
• v.2 no.1
• /
• pp.17-31
• /
• 1972

Light and electron microscopic studies were made to investigate the morphological changes in growing galls on the leaf of Lycium chinense MILL caused by Eriophyes kuko Kishda. The results are summarized as follows: 1. Light microscopy At the early stage of the invasion of E. kuko on the back side of the young leaf of L. chinense, the.epidermal cells become hypertrophic and develope a gall. As the gall grows, the cells of both palisade and spongy-layers become hypertrophic and these tissues are hard to be distinguished because of their irregular outgrowth. As the gall grows, the nuclei of the gall also become hypertrophic and larger than these of normal cells. 2. Electron microscopy Under electron microscopy the mitochondria, the golgi apparatus and the plastids of the advanced galls are degenerated and disintergrated and the cell walls become thicker than normal ones. The characteristic star bodies and the ring-form structures are found in the mature gall cells.

• Journal of Microbiology
• /
• v.38 no.4
• /
• pp.218-223
• /
• 2000

Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

• ALGAE
• /
• v.21 no.4
• /
• pp.407-416
• /
• 2006

Low molecular weight carbohydrates, phycobilin pigments and cell structure using light and transmission electron microscopy were used to describe a new genus of unicellular red algae, Erythrolobus coxiae (Porphyridiales, Porphyrideophyceae, Rhodophyta). The nucleus of Erythrolobus is located at the cell periphery and the pyrenoid, enclosed by a cytoplasmic starch sheath, is in the cell center. The pyrenoid matrix contains branched tubular thylakoids and four or more chloroplast lobes extend from the pyrenoid along the cell periphery. A peripheral encircling thylakoid is absent. The Golgi apparatus faces outward at the cell periphery and is always associated with a mitochondrion. Porphyridium and Flintiella, the other members of the Porphyrideophyceae, also lack a peripheral encircling thylakoid and have an ER-mitochondria-Golgi association. The low molecular weight carbohydrates digeneaside and floridoside are present, unlike both Porphyridium and Flintiella, which have only floridoside. The phycobilin pigments B-phycoerythrin, R-phycocyanin and allophycocyanin are present, similar to Porphyridium purpureum. The cells have a slow gliding motility without changing shape and do not require substrate contact. The ultrastructural features are unique to members of the Porphyrideophyceae and recent molecular analyses clearly establish the validity of this new red algal class and the genus Erythrolobus.

• The Plant Pathology Journal
• /
• v.29 no.1
• /
• pp.17-30
• /
• 2013

Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treat-ments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.

• Applied Biological Chemistry
• /
• v.32 no.1
• /
• pp.64-72
• /
• 1989

Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

• Animal cells and systems
• /
• v.12 no.2
• /
• pp.109-116
• /
• 2008

Among the four kinds of silk glands in the funnel-web spider Agelena limbata, the ampullate gland for dragline silk production is the most predominate one in both sexes, and is composed of three functional parts-excretory duct, storage ampulla and convoluted tail regions. Two pairs of major ampullate glands send secretory ductules to the anterior spinnerets, and another two pairs of minor ampullate glands supply the middle spinnerets. There are no apparent differences between the major and minor ampullate glands not only the external spigots but also their internal silk glands. However, the microstructure is very unique in this spider, because each gland has spherical shaped storage sac with twig-like branched tails. Nevertheless, the wall of the secretory region is similarly composed of a single layer of epithelial cells. The mature secretory silks in glandular epithelium are closely packed and accumulated as electron-opaque vesicles. Most of the secretory products which originated from the rough endoplasmic reticula(rER) are grown up by fusion with the surrounding small vesicles however, the Golgi complex does not seem to play an important role in this process of secretion.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.3
• /
• pp.398-407
• /
• 2020

Objective: The Yip1 domain family (YIPF) proteins were proposed to function in endoplasmic reticulum (ER) to Golgi transport and maintenance of the morphology of the Golgi, which were homologues of yeast Yip1p and Yif1p. YIPF3, the member 3 of YIPF family was a homolog of Yif1p. The aim of present study was to investigate the expression and regulation mechanism of porcine YIPF3. Methods: Quantitative realtime polymerase chain reaction (qPCR) was used to analyze porcine YIPF3 mRNA expression pattern in different tissues and pig kidney epithelial (PK15) cells stimulated by polyinosine-polycytidylic acid (poly [I:C]). Site-directed mutations combined with dual luciferase reporter assays and electrophoretic mobility shift assay (EMSA) were employed to reveal transcription regulation mechanism of porcine YIPF3. Results: Results showed that the mRNA of porcine YIPF3 (pYIPF3) was widely expressed with the highest levels in lymph and lung followed by spleen and liver, while weak in heart and skeletal muscle. Subcellular localization results indicated that it expressed in Golgi apparatus and plasma membranes. Upon stimulation with poly (I:C), the level of this gene was dramatically up-regulated in a time- and concentration-dependent manner. pYIPF3 core promoter region harbored three cis-acting elements which were bound by ETS proto-oncogene 2 (ETS2), zinc finger and BTB domain containing 4 (ZBTB4), and zinc finger and BTB domain containing 14 (ZBTB14), respectively. In which, ETS2 and ZBTB4 both promoted pYIPF3 transcription activity while ZBTB14 inhibited it, and these three transcription factors all played important regulation roles in tumorigenesis and apoptosis. Conclusion: The pYIPF3 mRNA expression was regulated by ETS2, ZBTB4, and ZBTB14, and its higher expression in immune organs might contribute to enhancing ER to Golgi transport of proteins, thus adapting to the immune response.