Purification of a terminal uridylyltransferase that acts as host factor in the in vitro poliovirus replicase reaction.

Andrews, Nancy C.; Baltimore, David

doi:10.1073/pnas.83.2.221

Cited by 63 publications

(39 citation statements)

References 39 publications

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“…This hypothesis is consistent with the observation that restriction of the growth of HAV in vitro appears to be at the level of RNA replication (D. A. Anderson et al, 1987 and the importance of host cell factors in poliovirus RNA synthesis (Morrow et al, 1985;Andrews & Baltimore, 1986). This proposed mechanism explains the association of decreasing virulence with gradual adaptation to growth in cell culture and implies that the determination of the molecular basis for attenuation may depend upon a better understanding of the mechanism for adaptation to growth of HAV in cell culture.…”

Section: Isupporting

confidence: 77%

Nucleotide Sequence of High-passage Hepatitis A Virus Strain HM175: Comparison with Wild-type and Cell Culture-adapted Strains

Ross

Anderson²,

Edwards³

et al. 1989

Journal of General Virology

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SUMMARYThe nucleotide sequence of cDNA from a high-passage, cell culture-adapted variant of hepatitis A virus strain HM175 was compared with the previously determined sequences of wild-type virus and two other cell culture-adapted variants. A total of 42 nucleotide changes were detected when the sequence was compared with wild-type virus. Five of these changes were common to all cell culture-adapted strains and a further two changes were shared by the strains that had experienced the greatest number of cell culture passages. The mutations were distributed throughout the genome coding for amino acid substitutions in regions 2B, 2C and 3D with silent changes in 1C and the 5' non-coding region. The possible relevance of these mutations to cell culture adaptation and attenuation is discussed.Hepatitis A virus (HAV) is a positive-stranded RNA virus which has been classified as an enterovirus within the Picornaviridae . Early attempts to isolate and passage HAV in cell culture were hampered by the absence of a cytopathic effect and the lack of a sensitive means of detecting viral antigen. Following the development of a sensitive immunofluorescence assay (Mathieson et al., 1977) for the detection of viral antigen, HAV was isolated in cell culture after several passages in non-human primates (Provost & Hilleman, 1979). Subsequently, primary isolation was achieved in a variety of cell cultures (Balayan et al., 1979; Frosner et al., 1979;Daemer et al., 1981). Serial passaging of HAV in these systems resulted in an increased yield of virus and a reduction in the time before maximum virus production (Provost & Hilleman, 1979; Frosner et al., 1979). Prolonged passage of HAV in cell culture has been shown to be accompanied by a decrease in virulence for experimentally infected animals and man (Provost et al., 1986;Purcell et al., 1984;Karron et al., 1988), but the molecular basis of these alterations in viral phenotype is not known.In an effort to define the genetic changes associated with adaptation to cell culture and attenuation of virulence, two groups have compared the complete nucleotide sequences of different cell culture-adapted variants of the HM 175 strain of HAV (Cohen et al., 1987 a;Jansen et al., 1988) with wild-type virus (Cohen et al., 1987c). Previously, we have described a partial nucleotide sequence comparison of HAV HM175 at passage level 59 (Ross et al., 1986;B. N. Anderson et al., 1988). The present paper compares the complete nucleotide sequence of this variant with both wild-type and the two other cell culture-adapted variants.Restriction enzyme fragments of the HM 175 cDNA clones depicted in Fig. 1 were subcloned into mp8 and mp9 derivatives of bacteriophage M13 and sequenced by the chain termination method (Sanger et al., 1977). All nucleotide changes from the wild-type HM175 sequence were confirmed by sequencing in both orientations in the M13 polylinker.The passage histories of the molecularly cloned variants ofHAV HM 175 are illustrated in Fig. 2. HM175m3 was derived from the initial human faecal...

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Section: Isupporting

confidence: 77%

Nucleotide Sequence of High-passage Hepatitis A Virus Strain HM175: Comparison with Wild-type and Cell Culture-adapted Strains

Ross

Anderson²,

Edwards³

et al. 1989

Journal of General Virology

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“…Rather, when analyzed with total RNA as substrate, a complex spectrum of larger labeled RNA molecules was observed with the HA-4 fraction (lane 10). This result is reminiscent of the former described unspecific cellular TUTase (Andrews and Baltimore 1986). Comparing lanes 4 and 9 of Figure 1A, the HA-3 fraction also contained significant amounts of the U6 snRNA-specific TUTase, however, slightly contaminated with the unspecific enzyme (lane 9).…”

Section: Purification Of U6-tutasementioning

confidence: 58%

“…That human U6-TUTase is one of only two RNA-uridylating enzymes of vertebrates that have been characterized in detail so far (Andrews and Baltimore 1986;Trippe et al 2003) and up to now the first one to be cloned. Based on its catalytical activity, the enzyme clearly belongs to the widespread and still growing superfamily of nucleotidyl transferases which are involved in a variety of metabolic pathways (Aravind and Koonin 1999).…”

Section: Discussionmentioning

confidence: 99%

“…A second, yet less well-characterized enzyme is involved in poliovirus replication. This cellular host factor provides the oligo(U) primer required for the initiation of RNA synthesis by the virus-encoded RNA-dependent RNA polymerase (Andrews and Baltimore 1986). On the average, this host enzyme adds five uridine residues, with no clear preference for virion RNA.…”

Section: Introductionmentioning

confidence: 99%

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Identification, cloning, and functional analysis of the human U6 snRNA-specific terminal uridylyl transferase

Trippe¹,

Guschina²,

Hoßbach³

et al. 2006

RNA

127

142

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Mammalian cells contain a highly specific terminal uridylyl transferase (TUTase) that exclusively accepts U6 snRNA as substrate. This enzyme, termed U6-TUTase, was purified from HeLa cell extracts and analyzed by microsequencing. All sequenced peptides matched a unique human cDNA coding for a previously unknown protein. Domain structure analysis revealed that the U6-TUTase also belongs to the well-characterized poly(A) polymerase protein superfamily. However, by amino acid sequence as well as RNA-binding motifs, human U6-TUTase is highly divergent from both the poly(A) polymerases and from the TUTases identified within the editing complexes of trypanosomes. After cloning, the recombinant U6-TUTase was expressed in HeLa cells. Analysis of its catalytical activity confirmed the identity of the cloned protein as U6-TUTase, exhibiting the same exclusive substrate specificity for U6 snRNA as the endogenous enzyme. That unique selectivity even excluded as substrate U6atac RNA, the functional homolog of the minor spliceosome. Finally, RNAi knockdown experiments revealed that U6-TUTase is essential for cell proliferation. Surprisingly, large amounts of the recombinant enzyme were found to accumulate within nucleoli.

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“…Andrews et al (1985) had suggested that this host factor could be a terminal uridylyltransferase (TUT), that is, an enzyme which adds U residues to the 3' terminus of an RNA molecule. In a further paper (Andrews & Baltimore, 1986a) it was then shown that TUT activity purified extensively with, and was inseparable from, the host factor. Preparations of TUT were capable of adding several U residues, around five in the conditions used, to the 3' poly(A) tract of poliovirus RNA.…”

Section: Picornavirusesmentioning

confidence: 99%

Some Highlights of Animal Virus Research in 1986

McGeoch

Howard

Desselberger

1987

Journal of General Virology

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In 1986 we published a review article entitled 'Some highlights of animal virus research in 1985', in which we attempted to summarize important and interesting research on animal viruses published in 1985 (McGeoch et al., 1986b). The limitations of the exercise were recognized: coverage was selective rather than systematic and, outside clearly pre-eminent subjects, was overtly influenced by personal scientific interests. In this article we now present a similar account of animal virology in 1986. Much the same ground rules have been retained. Selectivity remains of the first importance, and our treatment concerns work published in 1986 as far as possible, but with a little flexibility where necessary.As with 1985, animal virology in 1986 was dominated by work on acquired immune deficiency syndrome (AIDS) retroviruses. Outside this area, however, we were not able to identify any small set of truly outstanding advances, although work in many virus systems clearly made excellent progress. This evaluation posed problems for a review of the type undertaken, in particular for achieving a balanced selection of topics. With this background we also found it more necessary than before to address virus groups in separate sections (representing a step back from the previous, integrative stance). We have chosen to present aspects of research on, first, the AIDS viruses, and then in succession on hepatitis viruses, herpesviruses, poxviruses, papovaviruses, picornaviruses and negative strand viruses, and to finish with a section on virion structure. AIDSDuring 1986 the extent of the public health problem posed by AIDS continued to increase worldwide. Certainly, it was in this year that the scale, the immediacy and the longer term implications of this viral disease finally became a prominent part of public consciousness in Great Britain. Prospective cohort studies in Great Britain and in the U.S.A. (Goedert et al., 1986) demonstrated a high and increasing incidence of AIDS 3 to 5 years after infection with the AIDS virus. Also, in 1986, the very wide spread of infection in several Central African countries became much more clearly recognized (Mann et al., 1986a, b;Quinn et al., 1986). Basic research on the retrovirus responsible for AIDS continued to advance rapidly, but problems of treatment and prevention appear to be of a more intransigent nature. On a somewhat bathetic note, we register that the AIDS retrovirus has now been assigned the vernacular name 'human immunodeficiency virus' (HIV) (Coffin et al., 1986;). Here we recount aspects of virus genome structure and variation, gene function, and approaches to countering the virus.The wide variations in the genomes of HIV isolates were further examined. Alizon et al. (1986) determined the complete genomic sequences of two strains from Zaire, in Central Africa. These had biological properties indistinguishable from those of other HIV isolates, and their sequences showed the same layout of reading frames and other features, but sequence comparisons revealed a much greater degr...

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Purification of a terminal uridylyltransferase that acts as host factor in the in vitro poliovirus replicase reaction.

Cited by 63 publications

References 39 publications

Nucleotide Sequence of High-passage Hepatitis A Virus Strain HM175: Comparison with Wild-type and Cell Culture-adapted Strains

Nucleotide Sequence of High-passage Hepatitis A Virus Strain HM175: Comparison with Wild-type and Cell Culture-adapted Strains

Identification, cloning, and functional analysis of the human U6 snRNA-specific terminal uridylyl transferase

Some Highlights of Animal Virus Research in 1986

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