樊晓晖教授讲授 Virology Viruses: contain only Introduction: one kind of nucleic acidviron (DNA or RNA), 一、concept of virus and Obligate intracellular 二、relation with humanparasites beings Importance in medical microbiology Virion:intact、 Characteristics of viral mature,infectious diseases viral particle 3、Persistent disease 1、high infectivity 2、few effective drugs 4、Contributory factor in cancer How is virus found ? 1886, Mayer found tobacco disease 1892, Iwanowaski confirmed this disease was caused by tobacco mosaic virus. 1896, Beigerink named virus. Fig. Leaf of tobacco and EM of tobacco mosaic virus. SARS(2002)!!! Consequences of viral infections Suffering, followed by recovery Persistent disease Fatal disease Congenital disease Contributory factor in cancer Contributory factor in other diseases Some are asymptomatic! Viruses can be useful Vaccine development Gene therapy Tools to investigate host cells What are viruses? “A PIECE OF BAD NEWS WRAPPED UP IN A PROTEIN” What are viruses? Nucleic acid genome: DNA or RNA Protein coat Protection, entry Lipid envelope in some viruses Small (20-400nm) Obligate intracellular parasites GROWTH ON ARTIFICIAL MEDIA DIVISION BY BINARY FISSION CONTAINS BOTH RNA AND DNA CONTAINS RIBOSOMES CONTAINS MURAMIC ACID SENSITIVE TO ANTIBIOTICS BACTERIA + + + + + + MYCOPLASMA + + + + - + RICKETTSIA - + + + + + CHLAMYDIA - + + + - + VIRUSES - - - (-)* - - * The arenavirus family appears to ‘accidentally’ package ribosomes, but these appear to play no role in protein synthesis. Consequences Heavily parasitic on host cell No broad range antibiotics Host range May be insect/animal, insect/plant Do not cross eukaryote / prokaryote boundary May be wide or narrow Factors affecting host range Cell surface receptors Factors affecting host range o Cell surface receptors Availability of replication machinery Factors affecting host range Cell surface receptors Availability of replication machinery Ability to get out of cell and spread Factors affecting host range Cell surface receptors Availability of replication machinery Ability to get out of cell and spread Host anti-viral response Viral structure – some terminology Virus particle = virion Protein which coats the genome = capsid Capsid usually symmetrical Capsid + genome = nucleocapsid May have an envelope Characterization of virus (1)DNA/RNA,without cellular structure (2)Reproduce within cells (3)Reproductive fission----Replication C23 Size and shape of viruses Section 1 Size and shape •size: Range from 20nm to nm 300nm,most •measurement: less than150nm • shape: bricks……. most are spheres, some are rods, bullets, 第23章 病毒的基本性状 第二节 病毒的结构和化学组成 Section 2 Structure and composition of viruses 第23章 病毒的基本性状 第二节 第一节 病毒的结构和化学组成 病毒的大小与形态 structure: nucleic acid virion nucleocapsid capsid Non-envelope virion Some viruses have an envelope surrounding the nucleocapsid nucleic acid capsid envelope Enveloped virion 第23章 病毒的基本性状 第二节 第一节 病毒的结构和化学组成 病毒的大小与形态 core In the centre of virion Is composed of DNA or RNA functions:genome of viruse, provide information of propagation,genetics and variation for virus 第23章 病毒的基本性状 第二节 第一节 病毒的结构和化学组成 病毒的大小与形态 capsid The protein shell, or coat, that encloses the nucleic acid genome. be composed of capsomeres. composition:protein a. Protect the viral nucleic acid. functions b. Participate in the viral infection. c. Share the antigenicity 第23章 病毒的基本性状 第二节 病毒的结构和化学组成 第一节 病毒的大小与形态 Symmetry of Nucleocapsid According to the difference of number and arrangement of capsomeres : ①helical(Influenza Virus et al) ②Icosahedral( most of sphere virus) ③Complex(Poxvirus,Phage)。 第23章 病毒的基本性状 第二节 病毒的结构和化学组成 第一节 病毒的大小与形态 envelope A lipid-containing membrane that surrounds some viral particles It is acquired during viral maturation by a budding process through a cellular membrane, Viruses-encoded glycoproteins are exposed on the surface of the envelope. composition:lipid、protein and glucose. 第23章 病毒的基本性状 第二节第一节 病毒的结构和化学组成 病毒的大小与形态 Functions Antigenicity some viruses possess neuraminidase Infectivity Peplomere/spike associated with attachment and penetration of viruses Lipid protein cause fever. Resistance Damaged by drying, acid, detergent, and heat 第23章 病毒的基本性状 第二节 病毒的结构和化学组成 CHEMICAL COMPOSITION OF VIRUSES (一)、 Viral Nucleic Acid diversity intron Clone and exprssion (二)、 Viral Protein •capsid •Matrix and envelope Structural •Viral enzyme (polymerase) nonstructural •Inhibitory protein 第23章 第三节 病毒的培养与增殖 病毒的基本性状 Section 3 culture and propagation of viruses 一、 Culture of Viruses Grow in animal Viruses must live in cells by means of multiplication methods Embryonated eggs Organ and tissue culture Primary tissue culture cell lines Tissue culture include Secondary tissue culture cell lines Tumor or immortalized cell lines 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 propagation 二、 病毒的增殖 of viruses Unlike other microorganisms, viruses lack their enzymes that require for propagation, they must grow in susceptible and living cells. Mode of propagation——multiplication Virus must enter and replicate in living cells in order to “reproduce” themselves. Replicative cycle of viruses. 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 Replicative cycle of viruses This “growth cycle” involves specific attachment of virus, penetration and uncoating, nucleic acid transcription, protein synthesis, matureation and assembly of the virions and their subsequent release from the cell by budding or lysis Concept Step: Attachment and penetration Uncoating Biosythesis Maturation assembly and release 1、 Attachment and penetration : 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 Attachment: includes two phases ① Attachment is via ionic interactions which are temperature-independent ②Viral attachment protein recongnizes specific receptors on the cell surface Penetration Endocytoses Fussing Nucleocapsid directly penetrate 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 2、Uncoating Uncoating is usually achieved by cellular proteases “opening up” the capsid Happen before penetration: phage During the penetration Happen after penetration 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 can not detect virions in this phase 3、 BIOSYNTHESIS ECLIPSE phase : Basic step •Mode of biosythesis 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 Basic step of biosythesis Transcribe early mRNA Translate early protein such as polymerase and inhibitory protein. Replicate viral progeny nucleic acid Transcribe late mRNA and translate late protein (Structural protein)。 dsDNA Progeny dsDNA Early mRNA Late mRNA Early Pr (polymerase) Late Pr (Structural Pr) Herpesviruses Flow of events during the replication of dsDNA virus mRNA ssDNA ds+/-DNA Intermediate state Viral Pr New complementory DNA as template Replicate ssDNA Human Papovavirus Flow of events during the replication of ssDNA virus (+)ssRNA Being mRNA Translate early protein dsRNA (-)ssRNA intermediate state As template 1、Togavirus (+)ssRNA (late mRNA) progeny(+) ssRNA (non-segmented RNA) 2、Tumor virus Late protein ( Segmented RNA ) Flow of events during the replication of (+)ssRNA virus (-)ssRNA Protein (+)mRNA (structural and nonstructual) Viral polymerase ds(+/-)ssRNA (intermediate state) Progeny (-)ssRNA 1、Rhabdovirus ( non-segmented RNA ) 2、 Influenza virus (segmented RNA) Flow of events during the replication of (-)ssRNA virus Following host cell division (+)ssRNA Retro-polymerase (+/-)dsRNA:DNA RNase H hydrolyse (+)RNA HIV HTLV-I DNA polymerase (+/-)dsDNA Integrate host chromosome( pro-virus) mRNA Progeny virus Viral protein DNA HTLV-II Flow of events during the replication of retro-RNA virus Gene expression and genome replication Viral (+) mRNA 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 Methods for biosynthesis dsDNA viruses;ssDNA viruses; (+)ssRNA;(-)ssRNA viruses retro RNA viruses;dsRNA viruses. 第23章 病毒的基本性状 第三节 病毒的培养与增殖 二、 病毒的增殖 4、assembly、maturation and release Enveloped viruses : budding Non-enveloped viruses: eruption of cells. 第23章 病毒的基本性状 第三节 病毒的培养与增殖 三、Effect caused by viruses 1、 viruses propagate and cause cell lysing--Cytopathic effect: Non-enveloped viruses 2、 viruses propagate, cells do not lyse : Enveloped viruses 3、viral genes integrate intoTwo viruses infect one DNA viruses orinhibit retro-viruses cell, one virus chromosome of host cell: •Happen in the same strain of the viruses replication of 4、Interference of viruses : •Happen in different strain another virus of viruses •Dead viruses interfere lived viruses 第23章 病毒的基本性状 第三节 病毒的培养与增殖 三、 病毒增殖的细胞效应 Mechanism of interference Interferon indirectly inhibit replication of viruses Compete or destroy receptors on host cells, change the metabolism of host cells Block the mRNA translation of second virus. 第23章 病毒的基本性状 第三节 病毒的培养与增殖 四abnormal viral propagation defective virus : deficiency in some aspects of replication, but interfering the replication of normal viruses (腺病毒伴随病毒, 丁型肝炎病毒) 缺陷干扰颗粒(Defective interfering particles, DIP) 顿挫感染(abortive infection):When a virus infects a cell (or host), but cannot complete the full replication cycle ( not biosynthesize their components or not assemble virions.), i.e. a non-productive infection. 第23章 病毒的基本性状 Section 4 Physical and chemical effect on viruses 灭活(inactivation): lose infectivity; maintain antigenic properties ,ability to absorb RBC; physical factors:heat , cold, radiation; chemical factors:ether, detergents and formaldehyde. 第23章 病毒的基本性状 Section 5 classification of viruses 1、 Virus genome properties DNA viruses 2、 Virion morphology 3、 Virus protein proteries Retro viruses 4、 Physicochemical properties of the virion RNA viruses 第23章 病毒的基本性状 第五节 病毒的分类 卫星 (satellites)病毒,Dependovirus /Virusoids卫星病毒 分两类 RNA (拟病毒virusoid), help virus provides capsid RNA encodes capsid 类病毒(viroid): Viroids are small (200-400nt), circular RNA molecules with a rod-like secondary structure which possess no capsid or envelope which are associated with certain plant diseases. Their replication strategy like that of viruses - they are obligate intracellular parasites. 朊病毒(prion),亦称蛋白侵染子。 Prions are rather ill-defined infectious agents believed to consist of a single type of protein molecule with no nucleic acid component. Confusion arises from the fact that the prion protein & the gene which encodes it are also found in normal 'uninfected' cells. These agents are associated with diseases such as Creutzfeldt-Jakob disease in humans, scrapie in sheep & bovine spongiform encephalopathy (BSE) in cattle. Summary Viruses are the smallest infectious agents and contain nucleic acid (RNA or DNA )as their genomnome. They are not cells and are too small to be seen in the light microscope. They are basic life forms composed of a protein coat that surrounds genetic material, certain viruses are further enclosed by an external lipid bilayer membrane that surrounds the capsid and may contain glycoproteins. They are energy-less and float around until they come in contact with an appropriate cell. Replication of the genetic material occurs when the virus takes control of the host cell's synthetic machinery. Viruses contain all of the information, but usually do not possess all enzymes needs for replication. 第23章 病毒的基本性状 思考题 1 Describe the structure of virus 病 毒 的 结 构 .composition of those structures and their functions 各 结构有何化学组成?有何功能? 2 Describe the replicative cycle of viruses病毒的复制周期包括哪些阶段? 3 Definition: nucleocapsid 、 defective virus、interference Review questions 1. What are the two types of symmertry of viral capsid? 2. What are the functions of viral proteins? 3. What is the composition of the viral envelope, and how is it formed? 4. Which proteins of virus induce protective antibody? 5. What are the differences between viruses and prions?