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  1. Article ; Online: Complete laparoscopic choledochal cyst excision and hepaticojejunostomy with laparoscopic Roux-Y reconstruction using a 5-mm stapler: A case of a 2-month-old infant.

    Murakami, Masakazu / Kaji, Tatsuru / Nagano, Ayaka / Matsui, Mayu / Onishi, Shun / Yamada, Koji / Ieiri, Satoshi

    Asian journal of endoscopic surgery

    2021  Volume 14, Issue 4, Page(s) 824–827

    Abstract: ... Y reconstruction technically challenging. A 2-month-old girl diagnosed with a choledochal cyst ... Roux-Y reconstruction with a 5-mm stapler. After cyst excision, Roux-Y reconstruction was performed ...

    Abstract Laparoscopic choledochal cyst excision and hepaticojejunostomy for choledochal in children is performed only at particular institutions because of the technical difficulty. Although choledochal cyst excision and hepaticojejunostomy are feasible, the small working space of infants makes intraabdominal Roux-Y reconstruction technically challenging. A 2-month-old girl diagnosed with a choledochal cyst underwent complete laparoscopic choledochal cyst excision and hepaticojejunostomy, including intraabdominal Roux-Y reconstruction with a 5-mm stapler. After cyst excision, Roux-Y reconstruction was performed in the intraabdominal cavity. Jejunojejunostomy was performed in a side-to-side fashion using a 5-mm stapler, and the entry hole was closed laparoscopically by hand-sewing, along with the mesentery defect. All procedures were performed by laparoscopic surgery. The postoperative course was uneventful, and the patient showed a quick recovery. Using small-diameter instruments, completely laparoscopic procedures for choledochal cyst are feasible.
    MeSH term(s) Abdominal Cavity ; Anastomosis, Roux-en-Y ; Anastomosis, Surgical ; Choledochal Cyst/surgery ; Common Bile Duct ; Female ; Humans ; Infant ; Laparoscopy
    Language English
    Publishing date 2021-02-15
    Publishing country Japan
    Document type Case Reports ; Journal Article
    ZDB-ID 2503256-2
    ISSN 1758-5910 ; 1758-5902
    ISSN (online) 1758-5910
    ISSN 1758-5902
    DOI 10.1111/ases.12928
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Different types of interaction between PCNA and PIP boxes contribute to distinct cellular functions of Y-family DNA polymerases.

    Masuda, Yuji / Kanao, Rie / Kaji, Kentaro / Ohmori, Haruo / Hanaoka, Fumio / Masutani, Chikahide

    Nucleic acids research

    2015  Volume 43, Issue 16, Page(s) 7898–7910

    Abstract: Translesion DNA synthesis (TLS) by the Y-family DNA polymerases Polη, Polι and Polκ, mediated via ...

    Abstract Translesion DNA synthesis (TLS) by the Y-family DNA polymerases Polη, Polι and Polκ, mediated via interaction with proliferating cell nuclear antigen (PCNA), is a crucial pathway that protects human cells against DNA damage. We report that Polη has three PCNA-interacting protein (PIP) boxes (PIP1, 2, 3) that contribute differentially to two distinct functions, stimulation of DNA synthesis and promotion of PCNA ubiquitination. The latter function is strongly associated with formation of nuclear Polη foci, which co-localize with PCNA. We also show that Polκ has two functionally distinct PIP boxes, like Polη, whereas Polι has a single PIP box involved in stimulation of DNA synthesis. All three polymerases were additionally stimulated by mono-ubiquitinated PCNA in vitro. The three PIP boxes and a ubiquitin-binding zinc-finger of Polη exert redundant and additive effects in vivo via distinct molecular mechanisms. These findings provide an integrated picture of the orchestration of TLS polymerases.
    MeSH term(s) Amino Acid Motifs ; Cell Line ; DNA/biosynthesis ; DNA-Directed DNA Polymerase/chemistry ; DNA-Directed DNA Polymerase/genetics ; DNA-Directed DNA Polymerase/metabolism ; Humans ; Mutation ; Proliferating Cell Nuclear Antigen/metabolism ; Protein Interaction Domains and Motifs ; Ubiquitination
    Chemical Substances Proliferating Cell Nuclear Antigen ; DNA (9007-49-2) ; DNA polymerase iota (EC 2.7.7.-) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; POLK protein, human (EC 2.7.7.7) ; Rad30 protein (EC 2.7.7.7)
    Language English
    Publishing date 2015-09-18
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 186809-3
    ISSN 1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
    ISSN (online) 1362-4962 ; 1362-4954
    ISSN 0301-5610 ; 0305-1048
    DOI 10.1093/nar/gkv712
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Defects in the striatal neuropeptide Y system in X-linked dystonia-parkinsonism.

    Goto, Satoshi / Kawarai, Toshitaka / Morigaki, Ryoma / Okita, Shinya / Koizumi, Hidetaka / Nagahiro, Shinji / Munoz, Edwin L / Lee, Lillian V / Kaji, Ryuji

    Brain : a journal of neurology

    2013  Volume 136, Issue Pt 5, Page(s) 1555–1567

    Abstract: Neuropeptide Y is a novel bioactive substance that plays a role in the modulation of neurogenesis ... a post-mortem analysis to determine the striatal localization profile of neuropeptide Y in neurologically ... of neuropeptide Y-positive neurons and numerous nerve fibres labelled for neuropeptide Y in the striatum ...

    Abstract Neuropeptide Y is a novel bioactive substance that plays a role in the modulation of neurogenesis and neurotransmitter release, and thereby exerts a protective influence against neurodegeneration. Using a sensitive immunohistochemical method with a tyramide signal amplification protocol, we performed a post-mortem analysis to determine the striatal localization profile of neuropeptide Y in neurologically normal individuals and in patients with X-linked dystonia-parkinsonism, a major representative of the neurodegenerative diseases that primarily involve the striatum. All of the patients examined were genetically verified as having X-linked dystonia-parkinsonism. In normal individuals, we found a scattered distribution of neuropeptide Y-positive neurons and numerous nerve fibres labelled for neuropeptide Y in the striatum. Of particular interest was a differential localization of neuropeptide Y immunoreactivity in the striatal compartments, with a heightened density of neuropeptide Y labelling in the matrix compartment relative to the striosomes. In patients with X-linked dystonia-parkinsonism, we found a significant decrease in the number of neuropeptide Y-positive cells accompanied by a marked loss of their nerve fibres in the caudate nucleus and putamen. The patients with X-linked dystonia-parkinsonism also showed a lack of neuropeptide Y labelling in the subventricular zone, where a marked loss of progenitor cells that express proliferating cell nuclear antigen was found. Our results indicate a neostriatal defect of the neuropeptide Y system in patients with X-linked dystonia-parkinsonism, suggesting its possible implication in the mechanism by which a progressive loss of striatal neurons occurs in X-linked dystonia-parkinsonism.
    MeSH term(s) Adult ; Aged ; Corpus Striatum/metabolism ; Corpus Striatum/pathology ; Dystonic Disorders/genetics ; Dystonic Disorders/metabolism ; Dystonic Disorders/pathology ; Female ; Genetic Diseases, X-Linked/genetics ; Genetic Diseases, X-Linked/metabolism ; Genetic Diseases, X-Linked/pathology ; Humans ; Male ; Middle Aged ; Neuropeptide Y/biosynthesis ; Neuropeptide Y/genetics
    Chemical Substances Neuropeptide Y
    Language English
    Publishing date 2013-05
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80072-7
    ISSN 1460-2156 ; 0006-8950
    ISSN (online) 1460-2156
    ISSN 0006-8950
    DOI 10.1093/brain/awt084
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  4. Article ; Online: Ingestion of eicosapentaenoic acid in the early stage of social isolation reduces a fibroblast growth factor 21 resistant state independently of body weight in KKA(y) mice.

    Nonogaki, Katsunori / Yamazaki, Tomoe / Murakami, Mari / Kaji, Takao

    Biochemical and biophysical research communications

    2015  Volume 464, Issue 2, Page(s) 674–677

    Abstract: ... hypertriglyceridemia associated with decreases in plasma insulin and FGF21 levels in KKA(y) mice while having no ... periostin, which are involved in hepatic lipogenesis and hepatosteaotosis, in KKA(y) mice. On the other hand ... by suppressing Notch- and gp78-independent SEREBP1c and ChREBP pathways in KKA(y) mice. ...

    Abstract Fibroblast growth factor (FGF) 21 is a mediator of glucose and lipid metabolism. Although exogenous administration of FGF21 exerts beneficial effects on glucose and lipid metabolism, circulating FGF21 levels are elevated in ob/ob and db/db mice, diet-induced obese mice and obese human. Here we show that ingestion of eicosapentaenoic acid (EPA) for 6 days after individually-housing significantly suppressed the hyperglycemia and hypertriglyceridemia associated with decreases in plasma insulin and FGF21 levels in KKA(y) mice while having no effects on food intake, body weight or plasma active GLP-1 levels. The ingestion of EPA had no significant effects on the expression of FGF21 in the liver, epididymal white adipose tissue and skeletal muscle. Moreover, the ingestion of EPA significantly decreased the expression of hepatic peroxisome sterol regulatory element-binding protein (SREBP1c), carbohydrate response element-binding protein (ChREBP), stearoyl-CoA deaturase and periostin, which are involved in hepatic lipogenesis and hepatosteaotosis, in KKA(y) mice. On the other hand, the ingestion of EPA had no significant effects on expression of hepatic gp78, Notch, forkhead box protein O1 or glucose-6-phosphatase. These findings suggest that EPA ingestion in the early stage of social isolation suppresses hyperglycemia and hypertriglyceridemia associated with reduced FGF21 and insulin resistance without altering food intake and body weight, and that the EPA ingestion suppresses hepatic lipogenesis by suppressing Notch- and gp78-independent SEREBP1c and ChREBP pathways in KKA(y) mice.
    MeSH term(s) Animals ; Blood Glucose/analysis ; Body Weight ; Eicosapentaenoic Acid/administration & dosage ; Eicosapentaenoic Acid/pharmacology ; Feeding Behavior/drug effects ; Fibroblast Growth Factors/blood ; Fibroblast Growth Factors/metabolism ; Gene Expression Regulation/drug effects ; Glucagon-Like Peptide 1/blood ; Insulin/blood ; Male ; Mice ; Social Isolation ; Triglycerides/blood
    Chemical Substances Blood Glucose ; Insulin ; Triglycerides ; fibroblast growth factor 21 ; Fibroblast Growth Factors (62031-54-3) ; Glucagon-Like Peptide 1 (89750-14-1) ; Eicosapentaenoic Acid (AAN7QOV9EA)
    Language English
    Publishing date 2015-08-21
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 205723-2
    ISSN 1090-2104 ; 0006-291X ; 0006-291X
    ISSN (online) 1090-2104 ; 0006-291X
    ISSN 0006-291X
    DOI 10.1016/j.bbrc.2015.07.058
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  5. Article ; Online: Laparoscopic Roux-en-Y gastric bypass and sleeve gastrectomy achieve comparable weight loss at 1 year.

    Yaghoubian, Arezou / Tolan, Amy / Stabile, Bruce E / Kaji, Amy H / Belzberg, Gary / Mun, Edward / Zane, Robert

    The American surgeon

    2012  Volume 78, Issue 12, Page(s) 1325–1328

    Abstract: ... to those after laparoscopic Roux-en-Y gastric bypass (RYGB), many of these studies are limited by the small ...

    Abstract Laparoscopic sleeve gastrectomy has gained popularity as a weight loss surgical option for morbidly obese patients. Although initial studies have shown weight loss and comorbidity resolution comparable to those after laparoscopic Roux-en-Y gastric bypass (RYGB), many of these studies are limited by the small patient size. Thus, the purpose of this study was to compare the outcomes of laparoscopic sleeve gastrectomy and laparoscopic RYGB. A retrospective chart review of all morbidly obese patients who underwent laparoscopic RYGB or sleeve gastrectomy between 2007 and 2009 at an HMO hospital was conducted. Data points collected included age, gender, completion of a preoperative weight loss program, initial body mass index (BMI), pre- and postoperative weights, and presence of diabetes mellitus (DM), hypertension (HTN), osteoarthritis, obstructive sleep apnea, and gastroesophageal reflux disease (GERD). Outcomes measures included excess weight loss, resolution of comorbidities, postoperative complications, and mortality. A total of 345 laparoscopic RYGBs and 192 sleeve gastrectomies were performed. On average, the patients who received RYGB were younger (46 vs 48 years, P = 0.05) and had higher BMI (47 vs 43 kg/m(2), P < 0.0001). There was a higher incidence of DM in the RYGB group (32 vs 22%, P = 0.01), whereas the incidences of HTN and GERD were similar in both surgical groups. Ninety-three per cent of the patients who underwent RYGB and 90 per cent of the patients who underwent sleeve gastrectomy completed a preoperative weight loss program. The median length of hospital stay for both groups was 3 days. The complication rate in both groups was 9 per cent. The incidence of gastric leak was 1 per cent in both groups. There was only one mortality, which occurred in the RYGB group. The postoperative resolution of DM was comparable in both groups. The RYGB group had greater resolution of HTN (48 vs 34%, P = 0.03) and GERD (73 vs 34%, P < 0.0001). At 12 months, sleeve gastrectomy achieved superior excess weight loss compared with RYGB (72 vs 61%, P = 0.0015). After adjusting for age and BMI, the excess weight loss for RYGB and sleeve gastrectomy was similar at 12 months (t parameter estimate -0.06, P = 0.08). Laparoscopic RYGB and sleeve gastrectomy had comparable postoperative morbidity and mortality rates. At 1 year, sleeve gastrectomy achieved only slightly greater weight loss. The two operations are both legitimate standalone bariatric procedures and their applications need to be based on individual patient characteristics and needs.
    MeSH term(s) Adult ; Anastomosis, Roux-en-Y/methods ; Body Mass Index ; California ; Cohort Studies ; Female ; Follow-Up Studies ; Gastrectomy/methods ; Gastric Bypass/methods ; Humans ; Laparoscopy/adverse effects ; Laparoscopy/methods ; Length of Stay ; Male ; Middle Aged ; Obesity, Morbid/diagnosis ; Obesity, Morbid/surgery ; Postoperative Complications/physiopathology ; Retrospective Studies ; Risk Assessment ; Time Factors ; Treatment Outcome ; Weight Loss
    Language English
    Publishing date 2012-12
    Publishing country United States
    Document type Comparative Study ; Journal Article
    ZDB-ID 202465-2
    ISSN 1555-9823 ; 0003-1348
    ISSN (online) 1555-9823
    ISSN 0003-1348
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  6. Article ; Online: Origin of the novel chemoreceptor Aesthetasc "Y" in Ostracoda: morphogenetical thresholds and evolutionary innovation.

    Kaji, Tomonari / Tsukagoshi, Akira

    Evolution & development

    2008  Volume 10, Issue 2, Page(s) 228–240

    Abstract: ... the Aesthetasc "Y" and the "Grouped setae," were compared. Cypridoidea and Pontocypridoidea, belonging ... our deduction is that the Aesthetasc "Y" originated from "Grouped setae-like" organ in the Paleozoic ... corresponding to those of the Aesthetasc "Y" as a common constraint in the molting process of setae. We conclude ...

    Abstract The morphology and developmental processes of the two types of ostracod chemoreceptors, the Aesthetasc "Y" and the "Grouped setae," were compared. Cypridoidea and Pontocypridoidea, belonging to Cypridocopina, have a large baseball bat-like seta as an autapomorphic character on the second antenna, whereas most ostracod taxa with plesiomorphic characters bear "Grouped setae" consisting of multiple setae on the second antenna. Their budding positions, morphology, and ontogenetic changes were compared, and our deduction is that the Aesthetasc "Y" originated from "Grouped setae-like" organ in the Paleozoic. The morphogenetic processes in the molting period of these chemoreceptors were compared at the cellular level. The observations suggest that the "Grouped setae" are formed by hypodermal cells and share sheath cells corresponding to those of the Aesthetasc "Y" as a common constraint in the molting process of setae. We conclude that modification of the morphogenetic processes in the molting period of the "Grouped setae" gave rise to the Aesthetasc "Y" as a novel organ in the evolutionary pathway of the Ostracoda.
    MeSH term(s) Animals ; Biological Evolution ; Chemoreceptor Cells/growth & development ; Chemoreceptor Cells/ultrastructure ; Crustacea/anatomy & histology ; Crustacea/classification ; Crustacea/genetics ; Crustacea/growth & development ; Larva/growth & development ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Morphogenesis ; Phylogeny
    Language English
    Publishing date 2008-03
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2020288-X
    ISSN 1525-142X ; 1520-541X
    ISSN (online) 1525-142X
    ISSN 1520-541X
    DOI 10.1111/j.1525-142X.2008.00230.x
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: [Toward a "Choosing Wisely" Future in Medicine].

    Kaji, Yuki

    Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan

    2019  Volume 139, Issue 4, Page(s) 547–550

    Abstract: The "Choosing Wisely" campaign is an activity to promote conversations between patients and doctors about unnecessary examinations, treatments, or procedures. A "Top five list" published by a number of specialty societies consists of five evidence-based ... ...

    Abstract The "Choosing Wisely" campaign is an activity to promote conversations between patients and doctors about unnecessary examinations, treatments, or procedures. A "Top five list" published by a number of specialty societies consists of five evidence-based recommendations in their own fields. Choosing Wisely Japan was launched in 2016; the campaign soon became widespread among primary care physicians. To make wiser choices in prescriptions of any medicine, it is necessary to consider the balance between its efficacies, risks, and costs. The purpose of this campaign is not only to publish recommendations to reduce waste in health care resources but also to disseminate and implement the recommendations contents. To put them into practice, it is necessary to think about interprofessional and interdisciplinary approaches.
    MeSH term(s) Cost-Benefit Analysis ; Evidence-Based Practice ; Health Resources/economics ; Health Resources/statistics & numerical data ; Humans ; Inappropriate Prescribing/prevention & control ; Interdisciplinary Communication ; Intersectoral Collaboration ; Japan ; Medical Overuse/prevention & control ; Patient Acceptance of Health Care ; Physician-Patient Relations ; Physicians, Primary Care ; Practice Patterns, Physicians'/economics ; Prescriptions/statistics & numerical data ; Risk Assessment
    Language Japanese
    Publishing date 2019-03-28
    Publishing country Japan
    Document type Journal Article ; Review
    ZDB-ID 200514-1
    ISSN 1347-5231 ; 0031-6903 ; 0372-7750 ; 0919-2085 ; 0919-2131
    ISSN (online) 1347-5231
    ISSN 0031-6903 ; 0372-7750 ; 0919-2085 ; 0919-2131
    DOI 10.1248/yakushi.18-00181-1
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  8. Article: Effect of synthetic porcine neuropeptide Y (NPY) on splanchnic blood flows and exocrine pancreatic secretion in dogs.

    Sumi, S / Inoue, K / Kogire, M / Doi, R / Yun, M / Kaji, H / Hosotani, R / Fujimura, M / Uchida, K / Kiyama, S / Kitagawa, K / Yajima, H / Fujii, N / Tobe, T

    Digestive diseases and sciences

    2009  Volume 36, Issue 11, Page(s) 1523–1528

    Abstract: This study examined the effect of synthetic porcine neuropeptide Y on the splanchnic blood flows ... and the exocrine pancreatic secretion in dogs. Graded doses of neuropeptide Y (0.1-5 microg/kg ... of the blood flows in the superior mesenteric artery, the portal vein, and the pancreatic tissue. Neuropeptide Y at 5 ...

    Abstract This study examined the effect of synthetic porcine neuropeptide Y on the splanchnic blood flows and the exocrine pancreatic secretion in dogs. Graded doses of neuropeptide Y (0.1-5 microg/kg, intravenous) caused dose-dependent reduction of the secretin-stimulated exocrine pancreatic secretion and of the blood flows in the superior mesenteric artery, the portal vein, and the pancreatic tissue. Neuropeptide Y at 5 microg/kg reduced the blood flows to 45.9 +/- 13.3% (superior mesenteric artery), 63.0 +/- 10.5% (portal vein), and 77.9 +/- 4.8% (pancreatic tissue), respectively. This dose also reduced secretin-stimulated pancreatic juice volume and CCK-8 plus secretin-stimulated protein output to 65.2 +/- 9.3 and 63.3 +/- 14.0%, respectively. This study shows a potent vasoconstrictor effect of neuropeptide Y on splanchnic vessels. Neuropeptide Y also inhibited exocrine pancreatic secretion in a significant correlation with the reduction in pancreatic tissue blood flow, which suggests that reduction in the blood flow may be one of the possible mechanisms of the inhibitory action of neuropeptide Y on exocrine secretion.
    MeSH term(s) Animals ; Dogs ; Female ; Male ; Neuropeptide Y/pharmacology ; Pancreas, Exocrine/blood supply ; Pancreas, Exocrine/drug effects ; Pancreas, Exocrine/metabolism ; Regional Blood Flow/drug effects ; Splanchnic Circulation/drug effects ; Swine
    Chemical Substances Neuropeptide Y
    Language English
    Publishing date 2009-01-21
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 304250-9
    ISSN 1573-2568 ; 0163-2116
    ISSN (online) 1573-2568
    ISSN 0163-2116
    DOI 10.1007/BF01296392
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  9. Article: Origin of the novel chemoreceptor Aesthetasc "Y" in Ostracoda: morphogenetical thresholds and evolutionary innovation

    Kaji, Tomonari / Tsukagoshi, Akira

    Evolution & development. 2008 Mar-Apr., v. 10, no. 2

    2008  

    Abstract: ... the Aesthetasc "Y" and the "Grouped setae," were compared. Cypridoidea and Pontocypridoidea, belonging ... our deduction is that the Aesthetasc "Y" originated from "Grouped setae-like" organ in the Paleozoic ... corresponding to those of the Aesthetasc "Y" as a common constraint in the molting process of setae. We conclude ...

    Abstract The morphology and developmental processes of the two types of ostracod chemoreceptors, the Aesthetasc "Y" and the "Grouped setae," were compared. Cypridoidea and Pontocypridoidea, belonging to Cypridocopina, have a large baseball bat-like seta as an autapomorphic character on the second antenna, whereas most ostracod taxa with plesiomorphic characters bear "Grouped setae" consisting of multiple setae on the second antenna. Their budding positions, morphology, and ontogenetic changes were compared, and our deduction is that the Aesthetasc "Y" originated from "Grouped setae-like" organ in the Paleozoic. The morphogenetic processes in the molting period of these chemoreceptors were compared at the cellular level. The observations suggest that the "Grouped setae" are formed by hypodermal cells and share sheath cells corresponding to those of the Aesthetasc "Y" as a common constraint in the molting process of setae. We conclude that modification of the morphogenetic processes in the molting period of the "Grouped setae" gave rise to the Aesthetasc "Y" as a novel organ in the evolutionary pathway of the Ostracoda.
    Language English
    Dates of publication 2008-03
    Size p. 228-240.
    Publisher Blackwell Publishing Inc
    Publishing place Malden, USA
    Document type Article
    ZDB-ID 2020288-X
    ISSN 1525-142X ; 1520-541X
    ISSN (online) 1525-142X
    ISSN 1520-541X
    DOI 10.1111/j.1525-142X.2008.00230.x
    Database NAL-Catalogue (AGRICOLA)

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  10. Article ; Online: Role of ribosome recycling factor in natural termination and translational coupling as a ribosome releasing factor.

    Inokuchi, Yoshio / Quaglia, Fabio / Hirashima, Akikazu / Yamamoto, Yoshihiro / Kaji, Hideko / Kaji, Akira

    PloS one

    2023  Volume 18, Issue 2, Page(s) e0282091

    Abstract: The role of ribosome recycling factor (RRF) of E. coli was studied in vivo and in vitro. We used the translational coupling without the Shine-Dalgarno sequence of downstream ORF (d-ORF) as a model system of the RRF action in natural termination of ... ...

    Abstract The role of ribosome recycling factor (RRF) of E. coli was studied in vivo and in vitro. We used the translational coupling without the Shine-Dalgarno sequence of downstream ORF (d-ORF) as a model system of the RRF action in natural termination of protein synthesis. For the in vivo studies we used the translational coupling by the adjacent coat and lysis genes of RNA phage GA sharing the termination and initiation (UAAUG) and temperature sensitive RRF. The d-ORF translation was measured by the expression of the reporter lacZ gene connected to the 5'-terminal part of the lysis gene. The results showed that more ribosomes which finished upstream ORF (u-ORF) reading were used for downstream reading when RRF was inactivated. The in vitro translational coupling studies with 027mRNA having the junction sequence UAAUG with wild-type RRF were carried out with measuring amino acids incorporation. The results showed that ribosomes released by RRF read downstream from AUG of UAAUG. In the absence of RRF, ribosomes read downstream in frame with UAA. These in vivo and in vitro studies indicate that RRF releases ribosomes from mRNA at the termination codon of u-ORF. Furthermore, the non-dissociable ribosomes read downstream from AUG of UAAUG with RRF in vitro. This suggests that complete ribosomal splitting is not required for ribosome release by RRF in translational coupling. The data are consistent with the interpretation that RRF functions mostly as a ribosome releasing factor rather than ribosome splitting factor. Additionally, the in vivo studies showed that short (less than 5 codons) u-ORF inhibited d-ORF reading by ribosomes finishing u-ORF reading, suggesting that the termination process in short ORF is not similar to that in normal ORF. This means that all the preexisting studies on RRF with short mRNA may not represent what goes on in natural termination step.
    MeSH term(s) Escherichia coli/genetics ; Ribosomal Proteins/genetics ; Ribosomes/genetics ; Codon, Terminator ; Peptide Chain Termination, Translational/genetics
    Chemical Substances ribosome releasing factor ; Ribosomal Proteins ; Codon, Terminator
    Language English
    Publishing date 2023-02-24
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0282091
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