分枝杆菌噬菌体 D29的生物学特性研究及其抗耐药结核潜力的探讨

目的研究分枝杆菌噬菌体 D29的生物学特性,为 D29抗耐药结核治疗奠定基础.方法观察噬菌体电镜结构和噬菌斑形态;测定 D29最佳感染复数(MOI);一步生长实验;检测 pH值对 D29活力的影响;斑点法测定裂解谱;中和实验检测抗原性.结果 D29噬菌斑圆形透明,边界清楚;D29尾长129nm,最佳 MOI为10-4;D29感染宿主菌的潜伏期约为50min,裂解量为10;pH值对 D29存活率影响大,酸性环境不影响 D29裂解能力;D29能裂解分枝杆菌临床耐药株;D29K值为1069.50.结论 D29属于长尾噬菌体科(siphoviridae),裂解谱广,抗原性较高,具有抗耐药结核潜力     

Partial complementation of a DNA ligase I deficiency by DNA ligase III and its impact on cell survival and telomere stability in mammalian cells

DNA ligase I (LigI) plays a central role in the joining of strand interruptions during replication and repair. In our current study, we provide evidence that DNA ligase III (LigIII) and XRCC1, which form a complex that functions in single-strand break repair, are required for the proliferation of mammalian LigI-depleted cells. We show from our data that in cells with either dysfunctional LigI activity or depleted of this enzyme, both LigIII and XRCC1 are retained on the chromatin and accumulate at replication foci. We also demonstrate that the LigI and LigIII proteins cooperate to inhibit sister chromatid exchanges but that only LigI prevents telomere sister fusions. Taken together, these results suggest that in cells with dysfunctional LigI, LigIII contributes to the ligation of replication intermediates but not to the prevention of telomeric instability.     

Genetic analysis of ubiquitin-dependent protein degradation.

Selective degradation of cellular proteins serves to eliminate abnormal proteins and to mediate the turnover of certain short-lived proteins, many of which have regulatory functions. In eukaryotes a major pathway for selective protein degradation is ATP-dependent and is mediated by the ubiquitin system. This pathway involves substrate recognition by components of a ubiquitin-protein ligase system, covalent attachment of ubiquitin moieties to proteolytic substrates, and subsequent degradation of these conjugates by a multicatalytic protease complex. Recent genetic evidence suggests that the remarkable selectivity of this process is largely controlled at the level of substrate recognition by the ubiquitin ligase system. In Saccharomyces cerevisiae, ubiquitin-conjugating enzymes UBC1, UBC4 and UBC5 have been identified as key components of this highly conserved degradation pathway. Genetic analysis indicates that ubiquitin-dependent proteolysis is essential for cell viability and that UBC4 and UBC5 enzymes are essential components of the eukaryotic stress response.     

RNA fragments rich in G and A nucleotides obligatory for in vitro DNA replication of phages phi-X 174 and lambda]

Evidence was presented that in vitro conversion of single-stranded DNA of phage phi X 174 to the double-stranded replicative form by partially purified DNA-dependent DNA polymerase I requires a specific RNA fragment acting as primer (25-50 nucleotides). RNA fragments highly rich in nucleotides A and G were obtained by partial degradation of E. coli M 500 Sho-R ribosomal RNA with pancreatic ribonuclease. They become covalently bound to the newly synthesized DNA chain of the replicative form of phage phi X 174. These RNA fragments are also required for in vitro replication of lambda phage DNA.     

Genetic analysis of ubiquitin-dependent protein degradation

Selective degradation of cellular proteins serves to eliminate abnormal proteins and to mediate the turnover of certain short-lived proteins, many of which have regulatory functions. In eukaryotes a major pathway for selective protein degradation is ATP-dependent and is mediated by the ubiquitin system. This pathway involves substrate recognition by components of a ubiquitin-protein ligase system, covalent attachment of ubiquitin moieties to proteolytic substrates, and subsequent degradation of these conjugates by a multicatalytic protease complex. Recent genetic evidence suggests that the remarkable selectivity of this process is largely controlled at the level of substrate recognition by the ubiquitin ligase system. InSaccharomyces cerevisiae, ubiquitin-conjugating enzymes UBC1, UBC4 and UBC5 have been identified as key components of this highly conserved degradation pathway. Genetic analysis indicates that ubiquitin-dependent proteolysis is essential for cell viability and that UBC4 and UBC5 enzymes are essential components of the eukaryotic stress response.     

Inhibition of the infective activity of bacteriophage f2 by spermine

Résumé La spermine a un effet inhibiteur sur la reproduction du phage f₂. Cette inhibition dépend de la concentration de la spermine. Ne diminuant pas le taux du phage libre et n'ayant pas d'effet sur l'adsorption, la spermine réduit l'activité de l'ARN infectieuse et la production du phage dans les sphéroplastes infectés.     

Structure and morphogenesis of bacteriophage T4

Bacteriophage T4 is one of the most complex viruses. More than 40 different proteins form the mature virion, which consists of a protein shell encapsidating a 172-kbp double-stranded genomic DNA, a tail, and fibers, attached to the distal end of the tail. The fibers and the tail carry the host cell recognition sensors and are required for attachment of the phage to the cell surface. The tail also serves as a channel for delivery of the phage DNA from the head into the host cell cytoplasm. The tail is attached to the unique portal vertex of the head through which the phage DNA is packaged during head assembly. Similar to other phages, and also herpes viruses, the unique vertex is occupied by a dodecameric portal protein, which is involved in DNA packaging.Received 18 February 2003; received after revision 16 April 2003; accepted 9 May 2003     

Regulation of Parkin E3 ubiquitin ligase activity

Parkin is an E3 ubiquitin ligase mutated in autosomal recessive juvenile Parkinson's disease. In addition, it is a putative tumour suppressor, and has roles outside its enzymatic activity. It is critical for mitochondrial clearance through mitophagy, and is an essential protein in most eukaryotes. As such, it is a tightly controlled protein, regulated through an array of external interactions with multiple proteins, posttranslational modifications including phosphorylation and S-nitrosylation, and self-regulation through internal associations. In this review, we highlight some of the recent studies into Parkin regulation and discuss future challenges for gaining a full molecular understanding of the regulation of Parkin E3 ligase activity.     

Construction of derivatives of lambda phage carrying the lamB gene inserted downstream from the promotors of the lactose operon]

The expression of gene lamB, the structural gene for the lambda receptor in E. coli K-12, has been put under the control of the promoter of the lactose operon. This has been done by in vitro recombination using vectors which are derivatives of phage lambda.     

噬菌体在癌症治疗研究中的应用

噬菌体作为一类特殊病毒能影响癌症进程.最新研究发现部分噬菌体通过活化免疫应答,影响细胞因子的分泌,调节肿瘤微环境,有益于癌症治疗.噬菌体还可作为筛选具有诱导抗肿瘤效应的外源肽的平台.本文综述噬菌体在癌症治疗研究中的应用,寻找攻克恶性肿瘤新途径.     

Eintritt und Mechanismus phagenhemmender Wirkungen

Summary The described antiphage substance: bis-[[4-[4-(3-diethylamino-propylamino)-6-methyl-2-pyrimidyl-amino]-phenyl]]-sulphone tetrahydrochloride tetrahydrate produces its effect not through a direct phagocytic action but indirectly through an inhibition of the phage production. The effect is complete during the whole logarithmic growth phase as long as the phage concentration is kept under the limit, critical for bacteriolysis.     

Diversity and roles of (t)RNA ligases

The discovery of discontiguous tRNA genes triggered studies dissecting the process of tRNA splicing. As a result, we have gained detailed mechanistic knowledge on enzymatic removal of tRNA introns catalyzed by endonuclease and ligase proteins. In addition to the elucidation of tRNA processing, these studies facilitated the discovery of additional functions of RNA ligases such as RNA repair and non-conventional mRNA splicing events. Recently, the identification of a new type of RNA ligases in bacteria, archaea, and humans closed a long-standing gap in the field of tRNA processing. This review summarizes past and recent findings in the field of tRNA splicing with a focus on RNA ligation as it preferentially occurs in archaea and humans. In addition to providing an integrated view of the types and phyletic distribution of RNA ligase proteins known to date, this survey also aims at highlighting known and potential accessory biological functions of RNA ligases.     

Preliminary observations of a bacteriophage infecting Xenorhabdus luminescens (Enterobacteriaceae)

A bacteriophage infective to Xenorhabdus luminescens, a bacterial symbiont of heterorhabditid nematodes, was recovered from insects that supported poor nematode development. Plaque tests showed the phage particles to be infective only to primary and not secondary colonies of X. luminescens. The phage was not infective to X. nematophilus primaries or secondaries. The bacteriophage particles ranged 80-90 nm in length, with the head ranging from 40 to 50 nm in diameter. Restriction analysis was performed on isolated bacteriophage DNA. This first report of a bacteriophage from Xenorhabdus species has practical implications since it could be detrimental to cultures of Heterorhabditis nematodes that are being produced throughout the world for the biological control of insects.     

Preliminary observations of a bacteriophage infectingXenorhabdus luminescens (Enterobacteriaceae)

A bacteriophage infective toXenorhabdus luminescens, a bacterial symbiont of heterorhabditid nematodes, was recovered from insects that supported poor nematode development. Plaque tests showed the phage particles to be infective only to primary and not secondary colonies ofX. luminescens. The phage was not infective toX. nenatophilus primaries or secondaries. The bacteriophage particles ranged 80–90 nm in length, with the head ranging from 40 to 50 nm in diameter. Restriction analysis was performed on isolated bacteriophage DNA. This first report of a bacteriophage fromXenorhabdus species has pratical implications since it could be detrimental to cultures ofHeterorhabditis nematodes that are being produced throughout the world for the biological control of insects.     

Ube3a,the E3 ubiquitin ligase causing Angelman syndrome and linked to autism,regulates protein homeostasis through the proteasomal shuttle Rpn10

Ubiquitination, the covalent attachment of ubiquitin to a target protein, regulates most cellular processes and is involved in several neurological disorders. In particular, Angelman syndrome and one of the most common genomic forms of autism, dup15q, are caused respectively by lack of or excess of UBE3A, a ubiquitin E3 ligase. Its Drosophila orthologue, Ube3a, is also active during brain development. We have now devised a protocol to screen for substrates of this particular ubiquitin ligase. In a neuronal cell system, we find direct ubiquitination by Ube3a of three proteasome-related proteins Rpn10, Uch-L5, and CG8209, as well as of the ribosomal protein Rps10b. Only one of these, Rpn10, is targeted for degradation upon ubiquitination by Ube3a, indicating that degradation might not be the only effect of Ube3a on its substrates. Furthermore, we report the genetic interaction in vivo between Ube3a and the C-terminal part of Rpn10. Overexpression of these proteins leads to an enhanced accumulation of ubiquitinated proteins, further supporting the biochemical evidence of interaction obtained in neuronal cells.     

5-Methylangelicin: a new highly photosensitizing angular furocoumarin

5-Methylangelicin, a new highly photosensitizing angular furocoumarin, was studied in 2 different biological systems, the T2 phage and Ehrlich ascites tumor cells; in comparison with angelicin, the parent compound, it was several times more active.     

Visualization of DNA in various phages (T4, χ, T7, ϕ 29) by ethidium bromide epi-fluorescent microscopy

Summary DNA-containing areas in various phages (T4, , T7 and 29) could be observed at the light microscopic level using ethidium bromide epi-fluorescent microscopy. The fluorescent intensity per phage was in linear proportion to the DNA content in each phage.Acknowledgements. This work was supported in part by the grant No. 521708 and No. 511212 from the Japan Ministry of Education, Science and Culture.     

Spécificité de l'action antiphage de substances synthétiques

Summary The effect of synthetic antiphages has been studied against different types of bacteriophage in the same bacterium, in this caseEscherichia coli. Most substances are active only against typesT ₁, 3, 5, 7 (and our type 207) but not against types 2, 4, 6. Others show activity only against type 1 (and our type 207). The antiphage activity of a substance therefore seems to depend upon a specific relationship between its chemical structure and that of the phage type against which it is active.