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Role of the PEST sequence in the long-type GATA-6 DNA-binding protein expressed in human cancer cell lines

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Advances in Bioscience and Biotechnology 2012年第4期3卷 314-320页

作者： kanako Obayashi Kayoko Takada Kazuaki Ohashi Ayako kobayashi-Ohashi Masatomo Maeda Department of Immunobiology School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Molecular Biology School of Pharmacy Iwate Medical University Shiwa-Gun Japan Departments of Medical Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Laboratory of Biochemistry and Molecular Biology Graduate School of Pharmaceutical Sciences Osaka University Suita Japan

GATA-6 mRNA utilizes two Met-codons in frame as translational initiation codons in cultured mammalian cells. Deletion of the nucleotide sequence encoding the PEST sequence between the two initiation codons unusually reduced the protein molecular size on SDS-polyacrylamide gel-electrophoresis. The reduced molecular size is ascribed to the molecular property of GATA-6, since both amino-and carboxy-lterminal tags introduced into GATA-6 were detected on the gel. This PEST sequence seems to contribute to expansion of the long-type GATA-6 molecule. The long-type GATA-6 containing the PEST sequence exhibits more activation potential than that without this sequence, the latter’s activity being similar to that of the short-type GATA-6. We further demonstrated that human colon and lung cancer cell lines express both the long-type GATA-6 and the short-type GATA-6 in their nuclei.

关键词： DNA-Binding Protein GATA-6 Transcription Factor Leaky Ribosome Scanning PEST Sequence Gel Electrophoresis

Distant metastasis of hepatocellular carcinoma to Meckel’s cave and cranial nerves:A case report and review of literature

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World Journal of Hepatology 2021年第6期13卷 709-716页

作者： Soo Ki Kim Takako Fujii Ryouhei Komaki Hisato kobayashi Toyokazu Okuda Yumi Fujii Takanobu Hayakumo kanako Yuasa Masahiro Takami Aya Ohtani Yuka Saijo Yu-Ichiro Koma Soo Ryang Kim Department of Gastroenterology Kobe Asahi HospitalKobe 653-0801HyogoJapan Division of Neurology Kobe University Graduate School of MedicineKobe 650-0017HyogoJapan Department of Radiology Kobe Asahi HospitalKobe 653-0801HyogoJapan Department of Pharmacy Kobe Asahi HospitalKobe 653-0801HyogoJapan Department of Pathology Kobe University Graduate School of MedicineKobe 650-0017HyogoJapan

BACKGROUND Metastasis occurs as a late event in the natural history of hepatocellular carcinoma(HCC),and most patients die of liver failure attributed to the tumor supplanting the ***,the brain is a less common metastatic *** SUMMARY We describe a rare case of hepatitis C virus-related multiple HCC metastasizing to the cavernous sinus,Meckel’s cave,and the petrous bone involving multiple cranial nerves in an 82-year-old *** admission imaging studies including Gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging(MRI)revealed multiple HCC nodules in both right and left *** guided biopsy of the left lobe revealed moderately differentiated *** targeted therapy with Lenvatinib(8 mg/d for 94 d,per os)and Ramucirumab(340 mg/d and 320 mg/d,two times by intravenous injection)were administered for 4 mo,resulting in progression of the *** months after the start of molecular target therapy,the patient presented with symptoms of hyperalgesia of the right face and limited abduction of the right eye,indicating disturbances in the right trigeminal and abducens *** MRI disclosed a mass involving the cavernous sinus,Meckel’s cave and the petrous ***-enhanced MRI with gadolinium-chelated contrast medium revealed a well-defined mass with abnormal enhancement around the right cavernous sinus and the right Meckel’s *** The diagnosis of metastatic HCC to the cavernous sinus,Meckel’s cave,and the petrous bone was made based on neurological findings and imaging studies including MRI,but not on histological *** studies may provide insights into various methods for diagnosing HCC metastasizing to the craniospinal area.

关键词： Meckel’s cave Abducens nerve Trigeminal nerve Hepatocellular carcinoma Magnetic resonance imaging Case report

Increased Electrophoretic Mobility of Long-Type GATA-6 Transcription Factor upon Substitution of Its PEST Sequence

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Advances in Bioscience and Biotechnology 2014年第13期5卷 1032-1042页

作者： kanako Obayashi Kayoko Takada Kazuaki Ohashi Ayako Ohashi-kobayashi Mayumi Nakanishi-Matsui Makoto Araki Masatomo Maeda Laboratory of Biochemistry and Molecular Biology Graduate School of Pharmaceutical Sciences Osaka University Suita Japan Department of Medical Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Immunobiology School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Molecular Biology School of Pharmacy Iwate Medical University Shiwa-Gun Japan

The transcriptional factor GATA-6 gene produces two translational isoforms from a single mRNA through ribosomal leaky scanning. L-type GATA-6 has an extension of 146 amino acid residues at its amino terminus. In the extension, there is a unique PEST sequence (Glu31-Cys46), which is composed of an amino terminal Pro-rich segment and a carboxyl terminal Ser-cluster. Substitution of either half of the PEST sequence with Ala residues by cassette mutagenesis reduced the apparent molecular size of L-type GATA-6 on SDS-polyacrylamide gel-electrophoresis. However, the effect of substitution of the Pro-rich segment was much more significant;the mobility increase of the Pro-rich segment on the gel was 13% while that of the Ser-cluster was 8%. Substitution of each amino acid residue demonstrated that the effect of Pro substitution is greater than that of the Ser and Thr residues. Such increased mobility of L-type GATA-6 in the presence of a detergent may apparently correlate with the decrease in transcription activity in vivo as determined by means of luciferase reporter gene assay. The activity of ΔAla (with Ala residues instead of the PEST sequence) was reduced to one fifth of that of ΔA (with the PEST sequence). These results suggest that the PEST sequence of L-type GATA-6 does not function as a constitutive protein degradation signal, but rather plays structural and functional roles in the activation of gene expression on the GATA responsive promoter.

关键词： Cassette Mutagenesis Long-Type GATA-6 Mobility on Gel-Electrophoresis PEST Sequence Proline-Rich Segment Transcription Factor

Preparation of Chiral Hydroxy Esters Using Actinobacteria: Biocatalyst Activity of Marine-Derived <i>Micromonospora</i>and <i>Streptomyces</i>Strains

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Open Journal of Applied Sciences 2013年第1期3卷 116-122页

作者： Kohji Ishihara Aiko Fujita Akane Sakiyama Yuko kobayashi Kaoru Hori kanako Maruike Noriyoshi Masuoka Nobuyoshi Nakajima Hiroki Hamada Department of Life Science Okayama University of Science Okayama Japan Graduate School of Health and Welfare Okayama Prefectural University Soja Japan

To research the potential ability of marine-derived actinomycetes to act as biocatalysts, 8 Micromonospora strains and 5 Streptomyces strains were screened. Two recommended media (227 and 1076 media) and 2 modified media (1076-25% and P-1076-25% media) for liquid culture of these marine-derived actinomycetes were tested. As a result, 2 Micromonospora strains (Micromonospora sp. NBRC107096 and 107097) cultured with the 1076-25% medium and 2 Streptomyces strains (Streptomyces tateyamensis NBRC105048 and Streptomyces sp. NBRC105896) cultured with P-1076-25% medium showed a good growth. The stereoselective reduction of α-keto esters using these 4 actinomycetes was tested. As a result, it was found that these strains had a reducing activity toward various α-keto esters. The introduction of L-glutamate or sucrose as an additive remarkably increased the conversion ratios in the reduction of substrates by the Micromonospora strain. Furthermore, in the presence of L-alanine, Streptomyces tateyamensis NBRC105048 reduced ethyl pyruvate, ethyl 2-oxobutanoate, ethyl 2-oxopentanoate, ethyl 2-oxohexanoate, and ethyl 3-methyl-2-oxobutyrate to the corresponding α-hydroxy ester with a high conversion ratio and with excellent enantiomeric excess. Thus, we found that these marine-derived actinomycetes have great potential to be used as biocatalysts for stereoselective reduction of carbonyl compounds.

关键词： Marine Bacteria Micromonospora Streptomyces Biocatalyst Chiral Alcohol

Giant Mediastinal Neurofibroma in a Child with Neurofibromatosis Type I

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Surgical Science 2012年第12期3卷 564-567页

作者： Koichiro Uchida Tadao Okada Shohei Honda Hisayuki Miyagi Ryouji kobayashi Akihiro Iguchi kanako C. Kubota Taketomi Akinobu Department of Gastroenterological Surgery I Hokkaido University Graduate School of Medicine Sapporo Japan Department of Pediatrics Hokkaido University Graduate School of Medicine Sapporo Japan Department of Surgical Pathology Hokkaido University Hospital Sapporo Japan

Aim: There are a variety of malignant tumors related to neurofibromatosis type 1 (NF1). This report describes a rare pediatric NF1 case with an unresectable giant mediastinal tumor. Case: A 6-year-old girl with wheezing was admitted to our institution for the further evaluation of a right mediastinal mass on plain chest radiography. On examination, there were multiple café au lait spots mainly on the trunk, and a well-defined, immobile, painless mass was palpable on her neck. The mediastinal lesion was detected as nonuniform mass surrounding the aortic arch, pulmonary artery, and right main bronchus on the contrast-enhanced CT and MRI. Open biopsy was useful to rule out malignancy and revealed neurofibroma, and contributed to follow up and treatment. Discussion: Open biopsy was useful to rule out malignancy, such as malignant peripheral nerve sheath tumor, revealed neurofibroma, and also contributed to follow up and treatment. The authors report successful management by open biopsy and discuss several clinical points regarding mediastinal neurofibroma for NF1.

关键词： Mediastinal Tumor Neurofibromatosis Type I Neurofibroma Open Biopsy

Scoring criteria for determining the safety of liver resection for malignant liver tumors

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World Journal of Meta-Analysis 2019年第5期7卷 234-248页

作者： Kohei Harada Minoru Nagayama Yoshiya Ohashi Ayaka Chiba kanako Numasawa Makoto Meguro Yasutoshi Kimura Hiroshi Yamaguchi Masahiro kobayashi Koji Miyanishi Junji Kato Toru Mizuguchi Departments of Surgery Surgical Scienceand OncologySapporo Medical UniversitySapporoHokkaido 060-8556Japan Division of Radiology Sapporo Medical University HospitalSapporoHokkaido 060-8556Japan Sapporo Medical University Postgraduate School of Health Science and Medicine Sapporo Medical UniversitySapporoHokkaido 060-8556Japan Research and Education Center for Clinical Pharmacy Kitasato University School of PharmacyTokyo 108-8641Japan Department of Internal Medicine IV Sapporo Medical UniversitySapporoHokkaido 060-8556Japan Department of Nursing and Surgical Science Sapporo Medical UniversitySapporo 0608543Japan

BACKGROUND Liver resection has become safer as it has become less ***,the minimum residual liver volume(RLV)required to maintain homeostasis is ***,the formulae used to calculate standard liver volume(SLV)are *** To review previously reported SLV formulae and the methods used to evaluate the minimum RLV,and explore the association between liver volume and *** A systematic review of Medline,PubMed,and grey literature was *** in the retrieved articles were cross-checked manually to obtain further *** last search was conducted on January 20,*** developed an SLV formula using data for 86 consecutive patients who underwent hepatectomy at our institution between July 2009 and August *** Linear regression analysis revealed the following formula:SLV(mL)=822.7×body surface area(BSA)?183.2(R2=0.419 and R=0.644,P<0.001).We retrieved 25 studies relating to SLV formulae and 12 studies about the RLV required for safe liver *** the previously reported formulae included various coefficient and constant values,a simplified version of the SLV,the common SLV(cSLV),can be calculated as follows:cSLV(mL)=710 or 770×*** minimum RLV for normal and damaged livers ranged from 20%-40%and 30%-50%,*** Sapporo score indicated that the minimum RLV ranges from 35%-95%depending on liver *** We reviewed SLV formulae and the minimum RLV required for safe liver *** Sapporo score is the only liver function-based method for determining the minimum RLV.

关键词： Standard liver volume Residual liver volume Hepatectomy Mortality Liver failure Liver function