Host ANP32 family proteins are crucial for maintaining the activity of influenza RNA polymerase and play an important role in the cross-species transmission of influenza viruses.To date,the molecular properties of equ...Host ANP32 family proteins are crucial for maintaining the activity of influenza RNA polymerase and play an important role in the cross-species transmission of influenza viruses.To date,the molecular properties of equine ANP32(eqANP32)protein are poorly understood,particularly the mechanisms that affect equine influenza virus(EIV)RNA polymerase activity.Here,we found that there are six alternative splicing variants of equine ANP32A(eqANP32A)with different levels of expression.Further studies showed that these six splicing variants of eqANP32A supported the activity of EIV RNA polymerase to varying degrees,with the variant eqANP32A_X2 having the highest expression abundance and exhibiting the highest support of polymerase activity.Sequence analysis demonstrated that the differences in the N-Cap regions of the six splicing variants significantly affected their N-terminal conformation,but did not affect their ability to bind RNA polymerase.We also demonstrated that there is only one transcript of eqANP32B,and that this transcript showed only very low support to the EIV RNA polymerase.This functional defect in eqANP32B is caused by the sequence of the 110–259 amino acids at its Cterminus.Our results indicated that it is the eqANP32A_X2 protein that mainly determines the efficiency of the EIV replication in horses.In conclusion,our study parsed the molecular properties of eqANP32 family proteins and revealed the sequence features of eqANP32A and eqANP32B,suggesting for the first time that the N-Cap region of ANP32A protein also plays an important role in supporting the activity of the influenza virus polymerase.展开更多
To amplify the NS, NP, PB1, PB2 and PA internal genes of two equine H3N8 influenza A viruses isolated in Greece in 2003 and 2007 five primer pairs were designed. The derived sequences were analysed from a phylogenetic...To amplify the NS, NP, PB1, PB2 and PA internal genes of two equine H3N8 influenza A viruses isolated in Greece in 2003 and 2007 five primer pairs were designed. The derived sequences were analysed from a phylogenetic point of view and compared with the evolutionary patters of the HA and NA proteins. Comparison of nucleotide sequences of the five internal genes of the Greek strains showed high similarity (99.3% - 99.7%) to strains isolated from outbreaks in Europe and Asia during 2002-2008. A total of 11 amino acid substitutions of the surface protein NA and the RNP complex proteins were identified in the Greek strains compared to those of progenitor viruses circulating up to 2003. These substitutions were repeated in Chinese and Mongolian isolates from outbreaks in 2007-2008. Notably NS1 protein did not acquired amino acid substitutions and moreover, a stop codon introduced at position 220 was stably maintained in the Greek strains. Phylogenetic trees of the five internal genes did not show the same separation in clades. Greek strains classified them into the American sublineage (as for the PA) Florida clade II (as for the NP, NS1 and PB1) and among Chinese strains of 2007-2008 outbreaks (as for the PB2). Additionally, evolutionary profiles of these internal proteins, except PB2, indicated a parallel evolution fashion to the HA protein, suggesting the possible occurrence of genetic reassortment between H3N8 viruses of district evolutionary lineages. In conclusion, phylogenetic analysis of the internal genes reported in this study could establish a candidate framework for future scientific communications on the phylogenetic diversity and evolution of the equine influenza viruses.展开更多
Standard schedules for equine influenza vaccinations have been published in Finland. However, no knowledge about the actual number of vaccinations has been available. Based on previous preliminary studies in slaughter...Standard schedules for equine influenza vaccinations have been published in Finland. However, no knowledge about the actual number of vaccinations has been available. Based on previous preliminary studies in slaughterhouses and among horse owners, it was suspected that vaccination recommendations are not always followed. The aims were to investigate vaccination status against equine influenza among Finnish racehorses in regional competitions. Vaccination status was investigated： （1） by utilizing database vaccination information of show jumping and dressage horses and （2） by carrying out a survey among horse industry experts. A total of 400 horses were selected at the beginning of the Finnish competition season in 2012 and five experts （key persons） were invited to a repeated survey implemented by Delphi techniques. Overall, 50%-100% of horses were properly vaccinated depending on the source available. Thirty-four percent （34%） of the horses had the competition license but no vaccination markings in the database. Differences between types of competition （show jumping and dressage） and between Southern and Northern parts of Finland were found. Vaccinations of show jumping horses were registered more often than vaccinations of dressage horses. Ninety percent （90%） of unvaccinated horses in the database were eight years or older. It can be concluded that equine influenza vaccination registry coverage among racehorses in Finland is incomplete. This poses a risk of misunderstanding, and the growing responsibility of competition organizers.展开更多
文摘流感病毒(Influenza Virus)根据血凝素(HA)和神经氨酸酶(NA)两种表面抗原蛋白分为不同的亚型,马流感Ⅰ型(H7N7)和Ⅱ型(H3N8)是其中比较重要的两个亚型。本研究应用无特定病原体(SPF)鸡胚增殖马流感病毒A/马/京防/74-1(H7N7)毒株,TRIzol LS Reagent提取病毒RNA,RT-PCR扩增HA基因全片段,克隆到PMD18-T载体上,并进行了鉴定和序列测定。所获得的HA基因片段长1727 bp,编码563个氨基酸残基。根据推导的氨基酸序列进行预测,有7个潜在的糖基化位点和16个半胱氨酸残基,通过序列分析推断,A/马/京防/74-1(H7N7)株病毒的HA来源于禽类,是一株通过基因重排出现的重组病毒。
基金the National Natural Science Foundation of China to HL Zhang(32002275)Natural Science Foundation of Heilongjiang Province of China to HL Zhang(YQ2020C021).
文摘Host ANP32 family proteins are crucial for maintaining the activity of influenza RNA polymerase and play an important role in the cross-species transmission of influenza viruses.To date,the molecular properties of equine ANP32(eqANP32)protein are poorly understood,particularly the mechanisms that affect equine influenza virus(EIV)RNA polymerase activity.Here,we found that there are six alternative splicing variants of equine ANP32A(eqANP32A)with different levels of expression.Further studies showed that these six splicing variants of eqANP32A supported the activity of EIV RNA polymerase to varying degrees,with the variant eqANP32A_X2 having the highest expression abundance and exhibiting the highest support of polymerase activity.Sequence analysis demonstrated that the differences in the N-Cap regions of the six splicing variants significantly affected their N-terminal conformation,but did not affect their ability to bind RNA polymerase.We also demonstrated that there is only one transcript of eqANP32B,and that this transcript showed only very low support to the EIV RNA polymerase.This functional defect in eqANP32B is caused by the sequence of the 110–259 amino acids at its Cterminus.Our results indicated that it is the eqANP32A_X2 protein that mainly determines the efficiency of the EIV replication in horses.In conclusion,our study parsed the molecular properties of eqANP32 family proteins and revealed the sequence features of eqANP32A and eqANP32B,suggesting for the first time that the N-Cap region of ANP32A protein also plays an important role in supporting the activity of the influenza virus polymerase.
文摘To amplify the NS, NP, PB1, PB2 and PA internal genes of two equine H3N8 influenza A viruses isolated in Greece in 2003 and 2007 five primer pairs were designed. The derived sequences were analysed from a phylogenetic point of view and compared with the evolutionary patters of the HA and NA proteins. Comparison of nucleotide sequences of the five internal genes of the Greek strains showed high similarity (99.3% - 99.7%) to strains isolated from outbreaks in Europe and Asia during 2002-2008. A total of 11 amino acid substitutions of the surface protein NA and the RNP complex proteins were identified in the Greek strains compared to those of progenitor viruses circulating up to 2003. These substitutions were repeated in Chinese and Mongolian isolates from outbreaks in 2007-2008. Notably NS1 protein did not acquired amino acid substitutions and moreover, a stop codon introduced at position 220 was stably maintained in the Greek strains. Phylogenetic trees of the five internal genes did not show the same separation in clades. Greek strains classified them into the American sublineage (as for the PA) Florida clade II (as for the NP, NS1 and PB1) and among Chinese strains of 2007-2008 outbreaks (as for the PB2). Additionally, evolutionary profiles of these internal proteins, except PB2, indicated a parallel evolution fashion to the HA protein, suggesting the possible occurrence of genetic reassortment between H3N8 viruses of district evolutionary lineages. In conclusion, phylogenetic analysis of the internal genes reported in this study could establish a candidate framework for future scientific communications on the phylogenetic diversity and evolution of the equine influenza viruses.
文摘Standard schedules for equine influenza vaccinations have been published in Finland. However, no knowledge about the actual number of vaccinations has been available. Based on previous preliminary studies in slaughterhouses and among horse owners, it was suspected that vaccination recommendations are not always followed. The aims were to investigate vaccination status against equine influenza among Finnish racehorses in regional competitions. Vaccination status was investigated： （1） by utilizing database vaccination information of show jumping and dressage horses and （2） by carrying out a survey among horse industry experts. A total of 400 horses were selected at the beginning of the Finnish competition season in 2012 and five experts （key persons） were invited to a repeated survey implemented by Delphi techniques. Overall, 50%-100% of horses were properly vaccinated depending on the source available. Thirty-four percent （34%） of the horses had the competition license but no vaccination markings in the database. Differences between types of competition （show jumping and dressage） and between Southern and Northern parts of Finland were found. Vaccinations of show jumping horses were registered more often than vaccinations of dressage horses. Ninety percent （90%） of unvaccinated horses in the database were eight years or older. It can be concluded that equine influenza vaccination registry coverage among racehorses in Finland is incomplete. This poses a risk of misunderstanding, and the growing responsibility of competition organizers.
