Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells
The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restr...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2020-10, Vol.104 (19), p.8455-8466 |
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| creator | Yuan, Hong Li, Na Li, Pinghua Bai, Xingwen Sun, Pu Bao, Huifang Gong, Xiaohua Ma, Xueqing Cao, Yimei Li, Kun Fu, Yuanfang Zhang, Jing Li, Dong Chen, Yingli Zhang, Jie Lu, Zengjun Liu, Zaixin |
| description | The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, this initiation codon was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA). Fortunately, the modifications did not prevent viral viability but influenced replication characteristics of some FMDV mutants in a cell-specific manner, as was shown by the similar replication in BHK-21 cells and delayed growth kinetics in PK-15 cells. This attenuated phenotype of FMDV mutants in PK-15 cells was found to be correlated with reduced abilities to cleave eIF4GI and suppress interference (IFN) expression. As leader (L) protein was reported to be responsible for eIF4GI cleavage and inhibition of IFN expression, the in vivo L protein synthesis was examined during the infection of FMDV mutants. Our results showed that not only the total yield of L proteins was severely influenced but also the individual yield of L protein was seen to be affected, which implied that both the relative usage of the two initiation sites and overall translation efficiency were changed by Lb AUG modifications. In addition, the in vitro translation activity was also negatively regulated by Lb AUG mutations. Collectively, these findings suggested that the restricted replications of Lb AUG-modified FMDVs were related to the delayed eIF4GI cleavage and decreased ability to block IFN expression but were mainly determined by the inefficient translation initiation. FMDVs precisely with modifications of Lb AUG initiation codon may represent safer seed viruses for vaccine production.
Key points
• The polyprotein translation of FMDV initiates at two alternative initiation sites (Lab AUG and Lb AUG). In order to explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, the Lb initiation AUG was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA), and four FMDV mutants with Lb AUG modification were generated.
• We found that partial FMDV mutants grew almost as well as WT virus in BHK-21 cells, a typical cell line used for FMD vaccine production, but displayed impaired replication in IFN-competent PK-15 cells.
• The attenuation of mutant FMDVs in PK-15 cells was found to be correlated with delayed eIF4GI cleavage and decreased abili |
| doi_str_mv | 10.1007/s00253-020-10810-w |
| format | Article |
| fullrecord | <record><control><sourceid>gale_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7471169</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A634466941</galeid><sourcerecordid>A634466941</sourcerecordid><originalsourceid>FETCH-LOGICAL-c612t-7d960015db0b7e9010c27ff4ee607fd7a11a1ddce4af997746f3a56967546e483</originalsourceid><addsrcrecordid>eNp9kktv1DAUhSMEotPCH2CBIrGhCxe_YscbpKriUVEE4rG2PMn1jKvEntpOCyv-Op6mtB2EUBaOcr9znGOfqnpG8BHBWL5KGNOGIUwxIrglGF09qBaEM4qwIPxhtcBENkg2qt2r9lM6x5jQVojH1R6jLcVMskX162PonXWdyS74Otg6r6FO0AXf1zkan4Z54rzLbn5NLkMdIeXoupyuBRE2wz0PG0JGxvdoDFNe171LYBLUly5OqTjVnz8g0tQdDEN6Uj2yZkjw9GY9qL6_ffPt5D06-_Tu9OT4DHWC0Ixkr0T5_aZf4qUEhQnuqLSWAwgsbS8NIYb0fQfcWKWk5MIy0wglZMMF8JYdVK9n3820HKGAvqQb9Ca60cSfOhindyferfUqXGrJJSFCFYOXNwYxXEwlvR5d2kYwHsKUNOVMMNUoSQv64i_0PEzRl3hbSsmmROJ31MoMoJ23oezbbU31sWCcC6E4KdTRP6jy9DC6cktgXfm-IzjcERQmw4-8MlNK-vTrl12WzmwXQ0oR7O15EKy3FdNzxXSpmL6umL4qouf3T_JW8qdTBWAzkMrIryDexf-P7W-rbtxZ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2439756124</pqid></control><display><type>article</type><title>Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells</title><source>MEDLINE</source><source>SpringerNature Journals</source><creator>Yuan, Hong ; Li, Na ; Li, Pinghua ; Bai, Xingwen ; Sun, Pu ; Bao, Huifang ; Gong, Xiaohua ; Ma, Xueqing ; Cao, Yimei ; Li, Kun ; Fu, Yuanfang ; Zhang, Jing ; Li, Dong ; Chen, Yingli ; Zhang, Jie ; Lu, Zengjun ; Liu, Zaixin</creator><creatorcontrib>Yuan, Hong ; Li, Na ; Li, Pinghua ; Bai, Xingwen ; Sun, Pu ; Bao, Huifang ; Gong, Xiaohua ; Ma, Xueqing ; Cao, Yimei ; Li, Kun ; Fu, Yuanfang ; Zhang, Jing ; Li, Dong ; Chen, Yingli ; Zhang, Jie ; Lu, Zengjun ; Liu, Zaixin</creatorcontrib><description>The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, this initiation codon was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA). Fortunately, the modifications did not prevent viral viability but influenced replication characteristics of some FMDV mutants in a cell-specific manner, as was shown by the similar replication in BHK-21 cells and delayed growth kinetics in PK-15 cells. This attenuated phenotype of FMDV mutants in PK-15 cells was found to be correlated with reduced abilities to cleave eIF4GI and suppress interference (IFN) expression. As leader (L) protein was reported to be responsible for eIF4GI cleavage and inhibition of IFN expression, the in vivo L protein synthesis was examined during the infection of FMDV mutants. Our results showed that not only the total yield of L proteins was severely influenced but also the individual yield of L protein was seen to be affected, which implied that both the relative usage of the two initiation sites and overall translation efficiency were changed by Lb AUG modifications. In addition, the in vitro translation activity was also negatively regulated by Lb AUG mutations. Collectively, these findings suggested that the restricted replications of Lb AUG-modified FMDVs were related to the delayed eIF4GI cleavage and decreased ability to block IFN expression but were mainly determined by the inefficient translation initiation. FMDVs precisely with modifications of Lb AUG initiation codon may represent safer seed viruses for vaccine production.
Key points
• The polyprotein translation of FMDV initiates at two alternative initiation sites (Lab AUG and Lb AUG). In order to explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, the Lb initiation AUG was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA), and four FMDV mutants with Lb AUG modification were generated.
• We found that partial FMDV mutants grew almost as well as WT virus in BHK-21 cells, a typical cell line used for FMD vaccine production, but displayed impaired replication in IFN-competent PK-15 cells.
• The attenuation of mutant FMDVs in PK-15 cells was found to be correlated with delayed eIF4GI cleavage and decreased ability to block IFN expression.
• We proved that the attenuated phenotype of Lb AUG-modified FMDVs was mainly determined by the inefficient translation initiation, as demonstrated by the decrease of total yield of L proteins and individual production of L protein.
• We successfully generated genetically engineered FMDV with attenuated phenotype. The approach of precise engineering of FMDV with the modification of initiation codon provides a safe platform to produce inactivated antigen vaccines.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-020-10810-w</identifier><identifier>PMID: 32820373</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Animals ; Antigens ; Applied Microbial and Cell Physiology ; Attenuation ; Biomedical and Life Sciences ; Biotechnology ; Cell Line ; Cleavage ; Codon ; Codon, Initiator ; Codons ; Foot & mouth disease ; Foot-and-Mouth Disease ; Foot-and-Mouth Disease Virus - genetics ; Genetic engineering ; Genetic translation ; Genotype & phenotype ; Growth kinetics ; Health aspects ; Interferon ; L protein ; Li, Dong ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Mutants ; Mutation ; Phenotypes ; Protein biosynthesis ; Protein Processing, Post-Translational ; Protein sorting signals ; Protein synthesis ; Proteins ; Replication ; Replication initiation ; Translation ; Translation initiation ; Vaccines ; Virus Replication ; Viruses</subject><ispartof>Applied microbiology and biotechnology, 2020-10, Vol.104 (19), p.8455-8466</ispartof><rights>The Author(s) 2020</rights><rights>COPYRIGHT 2020 Springer</rights><rights>The Author(s) 2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c612t-7d960015db0b7e9010c27ff4ee607fd7a11a1ddce4af997746f3a56967546e483</citedby><cites>FETCH-LOGICAL-c612t-7d960015db0b7e9010c27ff4ee607fd7a11a1ddce4af997746f3a56967546e483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-020-10810-w$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-020-10810-w$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32820373$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yuan, Hong</creatorcontrib><creatorcontrib>Li, Na</creatorcontrib><creatorcontrib>Li, Pinghua</creatorcontrib><creatorcontrib>Bai, Xingwen</creatorcontrib><creatorcontrib>Sun, Pu</creatorcontrib><creatorcontrib>Bao, Huifang</creatorcontrib><creatorcontrib>Gong, Xiaohua</creatorcontrib><creatorcontrib>Ma, Xueqing</creatorcontrib><creatorcontrib>Cao, Yimei</creatorcontrib><creatorcontrib>Li, Kun</creatorcontrib><creatorcontrib>Fu, Yuanfang</creatorcontrib><creatorcontrib>Zhang, Jing</creatorcontrib><creatorcontrib>Li, Dong</creatorcontrib><creatorcontrib>Chen, Yingli</creatorcontrib><creatorcontrib>Zhang, Jie</creatorcontrib><creatorcontrib>Lu, Zengjun</creatorcontrib><creatorcontrib>Liu, Zaixin</creatorcontrib><title>Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, this initiation codon was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA). Fortunately, the modifications did not prevent viral viability but influenced replication characteristics of some FMDV mutants in a cell-specific manner, as was shown by the similar replication in BHK-21 cells and delayed growth kinetics in PK-15 cells. This attenuated phenotype of FMDV mutants in PK-15 cells was found to be correlated with reduced abilities to cleave eIF4GI and suppress interference (IFN) expression. As leader (L) protein was reported to be responsible for eIF4GI cleavage and inhibition of IFN expression, the in vivo L protein synthesis was examined during the infection of FMDV mutants. Our results showed that not only the total yield of L proteins was severely influenced but also the individual yield of L protein was seen to be affected, which implied that both the relative usage of the two initiation sites and overall translation efficiency were changed by Lb AUG modifications. In addition, the in vitro translation activity was also negatively regulated by Lb AUG mutations. Collectively, these findings suggested that the restricted replications of Lb AUG-modified FMDVs were related to the delayed eIF4GI cleavage and decreased ability to block IFN expression but were mainly determined by the inefficient translation initiation. FMDVs precisely with modifications of Lb AUG initiation codon may represent safer seed viruses for vaccine production.
Key points
• The polyprotein translation of FMDV initiates at two alternative initiation sites (Lab AUG and Lb AUG). In order to explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, the Lb initiation AUG was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA), and four FMDV mutants with Lb AUG modification were generated.
• We found that partial FMDV mutants grew almost as well as WT virus in BHK-21 cells, a typical cell line used for FMD vaccine production, but displayed impaired replication in IFN-competent PK-15 cells.
• The attenuation of mutant FMDVs in PK-15 cells was found to be correlated with delayed eIF4GI cleavage and decreased ability to block IFN expression.
• We proved that the attenuated phenotype of Lb AUG-modified FMDVs was mainly determined by the inefficient translation initiation, as demonstrated by the decrease of total yield of L proteins and individual production of L protein.
• We successfully generated genetically engineered FMDV with attenuated phenotype. The approach of precise engineering of FMDV with the modification of initiation codon provides a safe platform to produce inactivated antigen vaccines.</description><subject>Animals</subject><subject>Antigens</subject><subject>Applied Microbial and Cell Physiology</subject><subject>Attenuation</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Cell Line</subject><subject>Cleavage</subject><subject>Codon</subject><subject>Codon, Initiator</subject><subject>Codons</subject><subject>Foot & mouth disease</subject><subject>Foot-and-Mouth Disease</subject><subject>Foot-and-Mouth Disease Virus - genetics</subject><subject>Genetic engineering</subject><subject>Genetic translation</subject><subject>Genotype & phenotype</subject><subject>Growth kinetics</subject><subject>Health aspects</subject><subject>Interferon</subject><subject>L protein</subject><subject>Li, Dong</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Phenotypes</subject><subject>Protein biosynthesis</subject><subject>Protein Processing, Post-Translational</subject><subject>Protein sorting signals</subject><subject>Protein synthesis</subject><subject>Proteins</subject><subject>Replication</subject><subject>Replication initiation</subject><subject>Translation</subject><subject>Translation initiation</subject><subject>Vaccines</subject><subject>Virus Replication</subject><subject>Viruses</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kktv1DAUhSMEotPCH2CBIrGhCxe_YscbpKriUVEE4rG2PMn1jKvEntpOCyv-Op6mtB2EUBaOcr9znGOfqnpG8BHBWL5KGNOGIUwxIrglGF09qBaEM4qwIPxhtcBENkg2qt2r9lM6x5jQVojH1R6jLcVMskX162PonXWdyS74Otg6r6FO0AXf1zkan4Z54rzLbn5NLkMdIeXoupyuBRE2wz0PG0JGxvdoDFNe171LYBLUly5OqTjVnz8g0tQdDEN6Uj2yZkjw9GY9qL6_ffPt5D06-_Tu9OT4DHWC0Ixkr0T5_aZf4qUEhQnuqLSWAwgsbS8NIYb0fQfcWKWk5MIy0wglZMMF8JYdVK9n3820HKGAvqQb9Ca60cSfOhindyferfUqXGrJJSFCFYOXNwYxXEwlvR5d2kYwHsKUNOVMMNUoSQv64i_0PEzRl3hbSsmmROJ31MoMoJ23oezbbU31sWCcC6E4KdTRP6jy9DC6cktgXfm-IzjcERQmw4-8MlNK-vTrl12WzmwXQ0oR7O15EKy3FdNzxXSpmL6umL4qouf3T_JW8qdTBWAzkMrIryDexf-P7W-rbtxZ</recordid><startdate>20201001</startdate><enddate>20201001</enddate><creator>Yuan, Hong</creator><creator>Li, Na</creator><creator>Li, Pinghua</creator><creator>Bai, Xingwen</creator><creator>Sun, Pu</creator><creator>Bao, Huifang</creator><creator>Gong, Xiaohua</creator><creator>Ma, Xueqing</creator><creator>Cao, Yimei</creator><creator>Li, Kun</creator><creator>Fu, Yuanfang</creator><creator>Zhang, Jing</creator><creator>Li, Dong</creator><creator>Chen, Yingli</creator><creator>Zhang, Jie</creator><creator>Lu, Zengjun</creator><creator>Liu, Zaixin</creator><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature 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of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells</title><author>Yuan, Hong ; Li, Na ; Li, Pinghua ; Bai, Xingwen ; Sun, Pu ; Bao, Huifang ; Gong, Xiaohua ; Ma, Xueqing ; Cao, Yimei ; Li, Kun ; Fu, Yuanfang ; Zhang, Jing ; Li, Dong ; Chen, Yingli ; Zhang, Jie ; Lu, Zengjun ; Liu, Zaixin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c612t-7d960015db0b7e9010c27ff4ee607fd7a11a1ddce4af997746f3a56967546e483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Antigens</topic><topic>Applied Microbial and Cell Physiology</topic><topic>Attenuation</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Cell Line</topic><topic>Cleavage</topic><topic>Codon</topic><topic>Codon, Initiator</topic><topic>Codons</topic><topic>Foot & mouth disease</topic><topic>Foot-and-Mouth Disease</topic><topic>Foot-and-Mouth Disease Virus - genetics</topic><topic>Genetic engineering</topic><topic>Genetic translation</topic><topic>Genotype & phenotype</topic><topic>Growth kinetics</topic><topic>Health aspects</topic><topic>Interferon</topic><topic>L protein</topic><topic>Li, Dong</topic><topic>Life Sciences</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Mutants</topic><topic>Mutation</topic><topic>Phenotypes</topic><topic>Protein biosynthesis</topic><topic>Protein Processing, Post-Translational</topic><topic>Protein sorting signals</topic><topic>Protein synthesis</topic><topic>Proteins</topic><topic>Replication</topic><topic>Replication initiation</topic><topic>Translation</topic><topic>Translation initiation</topic><topic>Vaccines</topic><topic>Virus Replication</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yuan, Hong</creatorcontrib><creatorcontrib>Li, 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USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yuan, Hong</au><au>Li, Na</au><au>Li, Pinghua</au><au>Bai, Xingwen</au><au>Sun, Pu</au><au>Bao, Huifang</au><au>Gong, Xiaohua</au><au>Ma, Xueqing</au><au>Cao, Yimei</au><au>Li, Kun</au><au>Fu, Yuanfang</au><au>Zhang, Jing</au><au>Li, Dong</au><au>Chen, Yingli</au><au>Zhang, Jie</au><au>Lu, Zengjun</au><au>Liu, Zaixin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2020-10-01</date><risdate>2020</risdate><volume>104</volume><issue>19</issue><spage>8455</spage><epage>8466</epage><pages>8455-8466</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, this initiation codon was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA). Fortunately, the modifications did not prevent viral viability but influenced replication characteristics of some FMDV mutants in a cell-specific manner, as was shown by the similar replication in BHK-21 cells and delayed growth kinetics in PK-15 cells. This attenuated phenotype of FMDV mutants in PK-15 cells was found to be correlated with reduced abilities to cleave eIF4GI and suppress interference (IFN) expression. As leader (L) protein was reported to be responsible for eIF4GI cleavage and inhibition of IFN expression, the in vivo L protein synthesis was examined during the infection of FMDV mutants. Our results showed that not only the total yield of L proteins was severely influenced but also the individual yield of L protein was seen to be affected, which implied that both the relative usage of the two initiation sites and overall translation efficiency were changed by Lb AUG modifications. In addition, the in vitro translation activity was also negatively regulated by Lb AUG mutations. Collectively, these findings suggested that the restricted replications of Lb AUG-modified FMDVs were related to the delayed eIF4GI cleavage and decreased ability to block IFN expression but were mainly determined by the inefficient translation initiation. FMDVs precisely with modifications of Lb AUG initiation codon may represent safer seed viruses for vaccine production.
Key points
• The polyprotein translation of FMDV initiates at two alternative initiation sites (Lab AUG and Lb AUG). In order to explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, the Lb initiation AUG was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA), and four FMDV mutants with Lb AUG modification were generated.
• We found that partial FMDV mutants grew almost as well as WT virus in BHK-21 cells, a typical cell line used for FMD vaccine production, but displayed impaired replication in IFN-competent PK-15 cells.
• The attenuation of mutant FMDVs in PK-15 cells was found to be correlated with delayed eIF4GI cleavage and decreased ability to block IFN expression.
• We proved that the attenuated phenotype of Lb AUG-modified FMDVs was mainly determined by the inefficient translation initiation, as demonstrated by the decrease of total yield of L proteins and individual production of L protein.
• We successfully generated genetically engineered FMDV with attenuated phenotype. The approach of precise engineering of FMDV with the modification of initiation codon provides a safe platform to produce inactivated antigen vaccines.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>32820373</pmid><doi>10.1007/s00253-020-10810-w</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2020-10, Vol.104 (19), p.8455-8466 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7471169 |
| source | MEDLINE; SpringerNature Journals |
| subjects | Animals Antigens Applied Microbial and Cell Physiology Attenuation Biomedical and Life Sciences Biotechnology Cell Line Cleavage Codon Codon, Initiator Codons Foot & mouth disease Foot-and-Mouth Disease Foot-and-Mouth Disease Virus - genetics Genetic engineering Genetic translation Genotype & phenotype Growth kinetics Health aspects Interferon L protein Li, Dong Life Sciences Microbial Genetics and Genomics Microbiology Mutants Mutation Phenotypes Protein biosynthesis Protein Processing, Post-Translational Protein sorting signals Protein synthesis Proteins Replication Replication initiation Translation Translation initiation Vaccines Virus Replication Viruses |
| title | Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells |
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