{"created":"2023-06-20T13:20:50.224282+00:00","id":894,"links":{},"metadata":{"_buckets":{"deposit":"af3b9bd1-b901-41a0-8624-5133c31c6025"},"_deposit":{"created_by":1,"id":"894","owners":[1],"pid":{"revision_id":0,"type":"depid","value":"894"},"status":"published"},"_oai":{"id":"oai:ir.soken.ac.jp:00000894","sets":["2:430:20"]},"author_link":["9854","9853","9852"],"item_1_creator_2":{"attribute_name":"著者名","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"安田, 二朗"}],"nameIdentifiers":[{}]}]},"item_1_creator_3":{"attribute_name":"フリガナ","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"ヤスダ, ジロウ"}],"nameIdentifiers":[{}]}]},"item_1_date_granted_11":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"1994-03-24"}]},"item_1_degree_grantor_5":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_name":"総合研究大学院大学"}]}]},"item_1_degree_name_6":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（理学）"}]},"item_1_description_12":{"attribute_name":"要旨","attribute_value_mlt":[{"subitem_description":" Roles of viral NS and M proteins on the influenza virus growth were
examined. The NS proteins of influenza virus, NS1I and NS2, are encoded
by RNA segment 8. The NSI proitein is encoded by a colinear mRNA
transcript, whereas thc NS2 protein with the molecular weight of 14.2
kilodaltons (kDa) is synthesid after splicing of NSI mRNA. Up to now,
these NS proteins are believed to exist only in virus-infected cells. The NSl
protein, which localizes in nuclei of virus-infected cells, recognizes the cis-
acting sequence on NSI mRNA and controls its splicing to NS2 mRNA.
NS2 is also present mainly in the nuclei. At present, however, little is
known on the function of this protein. This study indicates that the NS2
protein, previously considered as one of the two nonstructural proteins (NS 1
and NS2), exists in virus particles as a structural component. By
immunochemical method, the number of NS2 molecules in a virus particle
was estimated to be 130-200 molecules. After solubilization of viral
enve]ope, NS2 was still associated with ribonucleoprotein (RNP) cores, but
was later dissociated from RNP upon removal of the membrane M1 protein.
A filter-binding assay in vitro indlicated direct protein-protein contact
between M1 ancl NS2. Following chemical cleavage of the M1 protein,
NS2 was found to bincl only a C-terminal fragment of M1. By an
immunoprecipitation method, NS2-M1 complexes were also detected in
virus-infected cel] lysates. These observations altogether indicate specific
molecu]ar nteraction between M1 and NS2, suggestlng that NS2 regulates
the function of M1 or vice versa.
The M gene of influenza viruses encodes 2 proteins, M1 and M2.
The M 1 protein is tightly associated with virions forming a matrix, which
associates with RNP at its internal surface but interacts with envelope at its
external surface. M1 interacts with both NP, thereby interferes with the
function of RNP-associated RNA polymerase, and NS2 in virions. In
virus-infected cells, M1 is involved in both early (uncoating and import of
RNP into infectecl nuc]ei) and late (assembly of virions during maturation
and export of RNP from nuclei into cytoplasm) stages of virus growth. On
the othcr hand, M2 forms an ion channel and is considered to control the
transport of hemagglutinin (HA). M2 may also control uncoating step to
release RNP in the early phase of virus infection. Genetic studies described
in this report suggested that one or both of the M proteins have a regulatory
role(s) of the rate of virus growth. Influenza virus A/WSN/33 forms large
plaques (>3mm diameter) on MDCK cells whereas A/Aichi/2/68 forms
on]y small plaques (<1mm diameter). Fast growing reassortants (AWM),
isolated by mixed infection of MDCK cells with these two virus strains in
the presence of anti-WSN antibodies, all carried the M gene from WSN.
On MDCK cells, these reassortants produced progeny viruses as rapidly as
did WSN, and the virus yield was as high as Aichi. Pulse-labeling
experiments at various times after virus infectlon showed that the reassortant
AWM started to synthesize viral proteins earlier than Aichi. To determine
which of the two M proteins, M1 or M2, is responsible for the fast rate of
virus growth, an attempt was made to make recombinant viruses possessing
the chimeric M gene between WSN and Aichi. For this purpose, I
employed a newly developed RNA-transfection method into helper virus-
infected cells. The ts-mutant derived from WSN, ts5 1 , carrying the ts lesion
only in thc M gene, were used as a helper virus to rescue the chimeric M
gene RNA. A transfectant virus carrying a chimeric M gene consisting of
WSN-MI and Aichi-M2, CWA20, was generated by using an improved
reverse genetics system. The CWA20 virus formed large-sized plaques,
indicating that thc M1 protein, but not the M2 protein, was responsible for
this rapid growth of WSN-type. Taken together, I conclude that the
reassortant viruses entry into growth cycle faster than the parent Aichi strain,
presumably due to rapid uncoating of the M1 protein from RNP cores. As a
result of rapid uncoating, the reassortant RNP should be transported into
host nuclei faster than Aichi RNP, ultimately leading to an early onset of
transcription of the viral genes.","subitem_description_type":"Other"}]},"item_1_description_18":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_1_description_7":{"attribute_name":"学位記番号","attribute_value_mlt":[{"subitem_description":"総研大甲第88号","subitem_description_type":"Other"}]},"item_1_select_14":{"attribute_name":"所蔵","attribute_value_mlt":[{"subitem_select_item":"有"}]},"item_1_select_8":{"attribute_name":"研究科","attribute_value_mlt":[{"subitem_select_item":"生命科学研究科"}]},"item_1_select_9":{"attribute_name":"専攻","attribute_value_mlt":[{"subitem_select_item":"18 遺伝学専攻"}]},"item_1_text_10":{"attribute_name":"学位授与年度","attribute_value_mlt":[{"subitem_text_value":"1993"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"YASUDA, Jiro","creatorNameLang":"en"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-17"}],"displaytype":"simple","filename":"甲88_要旨.pdf","filesize":[{"value":"284.7 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"要旨・審査要旨 / Abstract, Screening Result","url":"https://ir.soken.ac.jp/record/894/files/甲88_要旨.pdf"},"version_id":"fb46043a-beea-4713-b2af-deeee2cd2f67"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-17"}],"displaytype":"simple","filename":"甲88_本文.pdf","filesize":[{"value":"1.8 MB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"本文","url":"https://ir.soken.ac.jp/record/894/files/甲88_本文.pdf"},"version_id":"ee0a8772-d403-42ed-9cc2-8152cfdec2b1"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"thesis","resourceuri":"http://purl.org/coar/resource_type/c_46ec"}]},"item_title":"インフルエンザウイルスの増殖の制御機構：ウイルス蛋白質の役割","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"インフルエンザウイルスの増殖の制御機構：ウイルス蛋白質の役割"},{"subitem_title":"Control of growth and assembly of influenza virus: Role of viral proteins","subitem_title_language":"en"}]},"item_type_id":"1","owner":"1","path":["20"],"pubdate":{"attribute_name":"公開日","attribute_value":"2010-02-22"},"publish_date":"2010-02-22","publish_status":"0","recid":"894","relation_version_is_last":true,"title":["インフルエンザウイルスの増殖の制御機構：ウイルス蛋白質の役割"],"weko_creator_id":"1","weko_shared_id":1},"updated":"2023-06-20T14:43:14.595559+00:00"}