Bacteriophage T4 gene transcription studied by hybridization to cloned restriction fragments

Young, Elton T.; Mattson, T.; Selzer, Gerald; Houwe, Griet Van; Bolle, A.; Epstein, Richard H.

doi:10.1016/s0022-2836(80)80011-8

Cited by 60 publications

(30 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the phage-infected r n e + and m e -cells (Fig. 2), in the absence of rifampicin, the rate of incorporation of [3H]uracil into RNA falls initially, as observed previously (Young et al 1980), and remains constant for the rest of the infection. After the addition of rifampicin, there is a rapid reduction in the incorporation of label, and the extent of this reduction is the same in both strains.…”

Section: E Coli Rnase E Affects the Functional Stability Of T4 Mrnassupporting

confidence: 79%

See 1 more Smart Citation

Escherichia coli RNase E has a role in the decay of bacteriophage T4 mRNA.

Mudd¹,

Carpousis²,

Krisch³

1990

Genes Dev.

View full text Add to dashboard Cite

Bacteriophage T4 mRNAs are markedly stabilized, both chemically and functionally, in an Escherichia coli strain deficient in the RNA-processing endonuclease RNase E. The functional stability of total T4 messages increased 6-fold; we were unable to detect a T4 message whose functional stability was not increased. There was a 4-fold increase in the chemical stability of total T4 RNA. The degree of chemical stabilization of six specific T4 mRNAs examined varied from a maximum of 28-fold to a minimum of 1.5-fold. In the RNase Edeficient strain, several minutes delay and a slower rate of progeny production led to a reduction in final phage yield of -50%. Although the effect of the rne temperature-sensitive mutation could be indirect, the simplest interpretation of our results is that RNase E acts directly in the degradation of many T4 mRNAs.[Key Words: Bacteriophage T4; mRNA decay; mRNA processing; mRNA stability; ribonuclease El Received September 13, 1989; revised version accepted February 7, 1990.Escherichia coli RNase E is an endoribonuclease that has been shown to process 9S RNA to a precursor of 5S rRNA (Ghora and Apirion 1978) and also to process RNA1, the inhibitor of ColE1 plasmid replication (Tomcs~inyi and Apirion 1985) both in vitro and in vivo. RNase E did not appear to have a general role in mRNA decay in E. coli (Apirion and Gitelman 1980}, although the synthesis of some E. coli proteins was affected by the mutation (Gitelman and Apirion 1980). The enzyme is involved in the processing of mRNA from bacteriophage T4 genes 32 and 59 in vivo. Processing of these messages resulted in destabilization of the portion of the mRNA upstream of the cleavage site (Mudd et al. 1988;Carpousis et al. 1989) and thus may have a role in retroregulation (Schmeissner et al. 1984) of upstream gene expression. A comparison of the cleavage sites found in the noncoding RNAs and the T4 mRNAs revealed similarities in sequence at the cleavage sites and in the potential to form RNA secondary structure just downstream of the sites (Tomcs~inyi and Apirion 1985;Mudd et al. 1988;Carpousis et al. 1989).As yet, the nucleases that determine the rate of functional decay of total mRNA in E. coil or its phage have not been identified. A mutation in the E. coli ams gene was found to have a five-to sixfold effect on the chemical stability of total E. coli RNAs, but it did not affect functional stability significantly (Kuwano et al. 1977;Ono and Kuwano 1979). E. coli RNases III (see Portier et al. 1987), E (Mudd et al. 1988;Carpousis et al. 1989), and other as yet unidentified endonucleases (Cannistraro et al. 1986; Baga et al. 1988; Melefors and von Gabain 1988; Uzan et al. 1988) have been implicated in the decay of a few specific bacterial or phage mRNAs. In this paper we present evidence suggesting that E. coli RNase E has a major role in the functional and chemical decay of many bacteriophage T4 mRNAs. Results E. coli RNase E affects the functional stability of T4 mRNAsThe functional stability of mRNA can be estimated from the ability of t...

show abstract

Section: E Coli Rnase E Affects the Functional Stability Of T4 Mrnassupporting

confidence: 79%

“…DNA filters were prepared and hybridized, without formamide, by using 5 ~g of labeled RNA, as described (Young et al 1980). The amounts of DNA bound per whole filter were 16 ~g of calf thymus, 32 ~g of whole T4, and 3.2 p.g of plasmid.…”

Section: Pulse-labeling Of Proteins and Rna Isolationmentioning

confidence: 99%

Escherichia coli RNase E has a role in the decay of bacteriophage T4 mRNA.

Mudd¹,

Carpousis²,

Krisch³

1990

Genes Dev.

View full text Add to dashboard Cite

show abstract

“…4). The products synthesized on the wild type DNA and on the DNA with deletion saA4 were compared and the following observations were made: (i)the two strong deletion-specific bands E4 and F4 are produced in the case of the deletion; (ii) the two (24). Recent measurements of the size of in vivo early RNA transcripts (T. Young and R. C. Menard, in preparation) have shown that the gene 52 message is present in an RNA molecule about 3.0 kb long, which corresponds to our G and H transcripts.…”

Section: Resultsmentioning

confidence: 99%

Mapping of in vitro transcription units and identification of primary transcripts of the D region of bacteriophage T4

Goldfarb

Burger

1981

Nucl Acids Res

View full text Add to dashboard Cite

The D region of bacteriophage T4 is comprised of six closely linked genes which are situated between 161 kb and 165 kb on the T4 chromosome. We studied the transcription of these genes in vitro by using DNA templates derived from a series of deletion mutants in this region. The mixture of primary products made by Escherichia coli RNA polymerase were fractionated by gel electrophoresis into discrete RNA species. The results obtained together with the known map positions of the deletions allowed to identify four wild-type and several deletion-specific transcripts of the D region. The end points of these transcripts were approximately mapped. The results demonstrate that the D region has two promoters and two terminators, an organisation which is similar to the previously established organisation of the T4 tRNA gene cluster.

show abstract

“…Relative transcription rates were determined by using a filter binding assay essentially as described previously (7,46 Hybridizations were carried out in glass scintillation vials in a shaking air incubator. Each vial contained two filters with immobilized pUCCAT11 or pGATet plasmid DNA plus one control filter with vector DNA in 0.5 ml of hybridization solution (4x SSC, 50% formamide, 0.4% SDS) with various amounts of labeled RNA.…”

Section: Methodsmentioning

confidence: 99%

A novel transcriptional response by the cat gene during slow growth of Escherichia coli

Meyer

Schottel

1991

J Bacteriol

View full text Add to dashboard Cite

A novel response to growth rate was found with expression of the chloramphenicol acetyltransferase (cat) gene in Escherichia coli. The amount of cat mRNA relative to total RNA increased about 11-fold as growth rates decreased 5-to 6-fold, without an increase in translation. The accumulation of cat mRNA was in contrast to decreased cellular concentrations of total RNA, trxA, ompA, or 23S rRNA as the growth rate decreased and was not due to changes in gene dosage or mRNA stability. Stability of the cat mRNA does not appear to be regulated by growth rate. No significant change in either chemical or functional stability was observed within a five-to sixfold range of growth rates when chemostat-grown cells were used. However, cat mRNA stability was affected by growth medium composition. The half-life of cat mRNA decreased about threefold, with an approximate fourfold increase in generation time due to changes in growth medium. Transcriptional studies have indicated that accumulation of cat mRNA at slow growth rates is the result of a specific transcriptional response to changes in cellular generation times. We propose that increases in the cellular concentration of a specific message at slow growth rates may reflect an additional type of survival response in E. coli.Regulation of gene expression in Escherichia coli frequently involves an interplay of transcriptional and posttranscriptional events that can be influenced by several parameters of growth. Growth rate is one of the parameters that has been implicated as an important factor affecting gene transcription and the stability of particular mRNAs in E. coli. At the transcriptional level, growth rate has been shown to affect tRNA synthesis (17) and to be involved in the proposed negative feedback regulation of rRNA synthesis (22) where transcription decreases with decreasing growth rate. Changes in transcription of a particular gene can obviously impact gene expression.Even though growth rate does not apparently influence the stability of most transcripts (5,12,20,21,37), it has been proposed to regulate the stability of the ompA and chloramphenicol acetyltransferase (cat) mRNAs in E. coli (32,35). Changes in message stability can affect the levels of protein synthesized from that transcript as seen for ompA (18), the puf operon of Rhodobacter capsulatus (11), and the pap (2) and ars (36) operons of E. coli. Therefore, in addition to transcriptional regulation, changes in mRNA stability due to changes in growth rate could have a critical effect on synthesis of the corresponding protein. Additional evidence of growth rate effects on gene expression can be seen in the increasing or decreasing levels of protein synthesis of numerous but largely uncharacterized E. coli proteins (38).Our interest has been in the events that regulate the stability of mRNA and thus expression of the cat gene. Although the stability of cat mRNA is reported to decrease with decreasing growth rate, the conditions used to vary growth rate involved carbon source changes (35). Since variations...

show abstract

Bacteriophage T4 gene transcription studied by hybridization to cloned restriction fragments

Cited by 60 publications

References 37 publications

Escherichia coli RNase E has a role in the decay of bacteriophage T4 mRNA.

Escherichia coli RNase E has a role in the decay of bacteriophage T4 mRNA.

Mapping of in vitro transcription units and identification of primary transcripts of the D region of bacteriophage T4

A novel transcriptional response by the cat gene during slow growth of Escherichia coli

Contact Info

Product

Resources

About