Characterization of a new class of deletions of the D region of the bacteriophage T4 genome

Depew, Richard E.; Snopek, Thomas J.; Cozzarelli, Nicholas R.

doi:10.1016/0042-6822(75)90086-0

Cited by 59 publications

(27 citation statements)

References 16 publications

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“…Pnk and Rnl1 are dispensable for T4 growth on common E. coli laboratory strains but required on a rare host encoding the optional locus prr (pnk and rnl1 restriction) (Depew & Cozzarelli, 1974;Sirotkin et al, 1978;Runnels et al, 1982;Jabbar & Snyder, 1984). Mutating a minuscule T4 orf termed stp (suppressor of three-prime phosphatase) abrogates prr restriction (Depew & Cozzarelli, 1974;Depew et al, 1975;Chapman et al, 1988;Penner et al, 1995). These facts reinforced the notion that Pnk and Rnl1 cooperate in RNA nick repair.…”

Section: Prrc -A Potential Phage-excluding Tool Counteracted By Trna supporting

confidence: 65%

RloC: A Translation-Disabling tRNase Implicated in Phage Exclusion During Recovery from DNA Damage

Kaufmann¹,

Davidov²,

Steinfels-Kohn³

et al. 2011

DNA Repair

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Section: Prrc -A Potential Phage-excluding Tool Counteracted By Trna supporting

confidence: 65%

RloC: A Translation-Disabling tRNase Implicated in Phage Exclusion During Recovery from DNA Damage

Kaufmann¹,

Davidov²,

Steinfels-Kohn³

et al. 2011

DNA Repair

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“…It is also possible that the sun Y gene plays an essential role under different growth conditions, such as during anaerobiosis or growth in an alternative host, such as Shigella dysenteriae (23). Preliminary screening of the ability of sun Y mutants to grow on a number of clinical isolates of E. coli that are known to restrict phage carrying mutations in other "nonessential" genes (8,9) has failed to identify a host that is restrictive for sun Y mutants.…”

Section: Methodsmentioning

confidence: 99%

“…Escherichia coli BE (supo) (3) and T4 deletion phages SaA9 and rNB2226 were provided by L. Gold. SaA9 contains a 2.4-kb deletion in the D region extending from ac through Dl (9), while rNB2226 contains an approximately 6-kb deletion spanning the rII region (1). E. coli XA101 [F-ara A(lac-pro) nalA supD metB argE(Am) rifthi] was from E. Eisenstadt.…”

Section: Methodsmentioning

confidence: 99%

The product of the split sunY gene of bacteriophage T4 is a processed protein

Zeeh

Shub

1991

J Bacteriol

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The sunY gene of bacteriophage T4 contains a self-splicing group I intron. The ligated exons encode an open reading frame of 605 anmo acids, whose inferred molecular mass is 68 kDa. However, none of the proteins made following T4 infection have been assigned to the sunY gene, and no mutations have been mapped to this locus. We show here that the primary product of the sunY gene is a protein with an apparent molecular mass of 64 kDa, which is processed to a protein approximately 4 kDa smaller. Unlike most other processed T4 proteins, cleavage occurs independently of both the T4 processing protease, the product of gene 21, and late phage protein synthesis. Insertional mutagenesis demonstrated that the sunY protein is not necessary for normal T4 growth under the conditions tested.The T4 sun Y gene is one of four bacteriophage genes that have been identified as containing self-splicing group I introns (7,12,30,32 deletion phages SaA9 and rNB2226 were provided by L. Gold. SaA9 contains a 2.4-kb deletion in the D region extending from ac through Dl (9), while rNB2226 contains an approximately 6-kb deletion spanning the rII region (1). E. coli XA101 [F-ara A(lac-pro) nalA supD metB argE(Am) rifthi] was from E. Eisenstadt. T4 alt (ac-q) (15) phage were from C. Goff, and amber mutants in T4 genes 21 (amHll) and 23 (amE322-amN12l) were from the collection of L. Black. J. Karam provided T4 gene 55 amber mutant amBL292 (3). T4 deletions farPl3, farP23, tk2, and tk25 (6) were from D. Hall, and deletion A9A (21), isolated by D.Parma, was from J. Wiberg.Descriptions of all relevant plasmids are given in intron (see Fig. 1). The rIIB-lacZ gene fusion of pAHZ2 was transferred to the sun Y gene of T4 rNB2226 by homologous recombination during infection of E. coli XA101 harboring the plasmid. The new phage was designated rNB2226, sun Y[rIIBlacAHZ2].A deletion within the sun Y gene was created in plasmid pAAX3 (Table 1) The sun Y deletion was introduced into phage, with or without a gene 55 amber mutation (amBL292), by recombination between plasmid DNA bearing the sun Y deletion and homologous sequences in the phage. The recipient phage used in this experiment contained a fusion of the sun Y intron open reading frame (ORF) with lacZ (13). Recombinant phage (containing the deletion) were identified by loss of P-galactosidase and were designated sun YA3 or 55am, sunYA3.Transfer of plasmid sequences to T4. Plasmid-containing strains of E. coli were grown in M9S medium (3) supplemented with 50 ,ug of ampicillin per ml to a density of 2 x 10R cells per ml and infected at a multiplicity of 1. After 5 min at 30°C, the cultures were diluted 100-fold into M9S medium and incubated for 1 to 2 h. Chloroform was added to lyse cells remaining in the culture. The progeny were plated and screened for their ability to form blue plaques on plates containing 0.04% 5-bromo-4-chloro-3-indolyl-,3-D-galactopyranoside (X-Gal) with a lac deletion lawn (E. coli XA101) (or for the loss of the ability to form blue plaques in the case of the sun Y internal del...

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“…To study their in vivo expression, each of these constructs was transferred into the phage genome via recombination between infecting phage DNA and homologous plasmid sequences flanking the fusions (Singer et al 1981;Casna and Shub 1982). Cells containing the fusion plasmids were infected with sah9, a phage containing a deletion of 2400 bp in the region between rIIB and gene 52 (Depew et al 1975). Use of a sizable deletion ensured that the T4 genome would be able to accommodate a 3-kb insert while still maintaining sufficient terminal redundancy for phage viability (Shub and Casna 1985).…”

Section: Expression Of the Intron Orfsmentioning

confidence: 99%

Genes within genes: independent expression of phage T4 intron open reading frames and the genes in which they reside.

Gott¹,

Zeeh²,

Bell-Pedersen³

et al. 1988

Genes & Development

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The td, ttrdB, and sunY introns of bacteriophage T4 each contain a long open reading frame (ORF). These ORFs are preceded by functional T4 late promoters and, in the case of the nrdB intron ORF, a functional middle promoter. Expression of phage-encoded intron ORF-IacZ fusions indicates that these T4 genes are highly regulated. The lack of translation of these ORFs from the early pre-mRNAs can be accounted for by the presence of secondary structures that are absent from the late RNAs. Because translation of the intron ORFs could disrupt core structural elements required for pre-mRNA splicing, such regulation may be necessary to allow expression of the genes in which they reside.

show abstract

Characterization of a new class of deletions of the D region of the bacteriophage T4 genome

Cited by 59 publications

References 16 publications

RloC: A Translation-Disabling tRNase Implicated in Phage Exclusion During Recovery from DNA Damage

RloC: A Translation-Disabling tRNase Implicated in Phage Exclusion During Recovery from DNA Damage

The product of the split sunY gene of bacteriophage T4 is a processed protein

Genes within genes: independent expression of phage T4 intron open reading frames and the genes in which they reside.

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