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Identification and sequence determination of the host factor gene for bacteriophage Q beta.

ABSTRACT: The host factor (HF-I) required for phage Q beta RNA-directed synthesis of complementary minus-strand RNA was purified to homogeneity from phage-infected Escherichia coli cells. The hfq gene encoding HF-I was cloned using synthetic probes designed based on the partial amino acid sequence of HF-I, and mapped at 94.8 min on the E. coli chromosome downstream of the miaA gene involved in 2-methylthio-N6-(isopentyl)-adenosine (ms2i6A) tRNA modification. Sequence determination of the cloned hfq gene indicated that HF-I is a small protein of Mr 11,166 consisting of 102 amino acid residues.

SUBMITTER: Kajitani M

PROVIDER: S-EPMC333781 | biostudies-other | 1991 Mar

REPOSITORIES: biostudies-other

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Identification and sequence determination of the host factor gene for bacteriophage Q beta.

Kajitani M M Ishihama A A

Nucleic acids research 19910301 5

The host factor (HF-I) required for phage Q beta RNA-directed synthesis of complementary minus-strand RNA was purified to homogeneity from phage-infected Escherichia coli cells. The hfq gene encoding HF-I was cloned using synthetic probes designed based on the partial amino acid sequence of HF-I, and mapped at 94.8 min on the E. coli chromosome downstream of the miaA gene involved in 2-methylthio-N6-(isopentyl)-adenosine (ms2i6A) tRNA modification. Sequence determination of the cloned hfq gene i ...[more]

PMID: 2020545

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Cloning and sequencing of hfq (host factor required for synthesis of bacteriophage Q beta RNA) gene of Salmonella Typhimurium isolated from poultry.

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Bacteroides thetaiotaomicron-Infecting Bacteriophage Isolates Inform Sequence-Based Host Range Predictions.

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2014-08-01 | E-GEOD-53348 | biostudies-arrayexpress

Determination and Inference of Eukaryotic Transcription Factor Sequence Specificity

2014-08-01 | GSE53348 | GEO

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Project description:Successful completion of the bacteriophage P2 lytic cycle requires phage-induced lysis of its Escherichia coli host, a process that is poorly understood. Genetic analysis of lysis-deficient mutants defined a single locus, gene K, which lies within the largest late transcription unit of P2 and maps between head gene L and tail gene R. We determined and analyzed the DNA sequence of a ca. 2.1-kb EcoRV fragment that spans the entire region from L to R, thus completing the sequence of this operon. This region contains all of the functions necessary for host cell lysis. Sequence analysis revealed five open reading frames, initially designated orf19 through orf23. All of the existing lysis mutants--ts60, am12, am76, and am218--were located in orf21, which must therefore correspond to gene K. The K gene product has extensive amino acid sequence similarity to the product of gene R of bacteriophage lambda, and its exhibits endolysin function. Site-directed mutagenesis and reverse genetics were used to create P2 amber mutants in each of the four other newly identified open reading frames. Both orf19 (gene X) and orf20 (gene Y) encode essential functions, whereas orf22 (lysA) and orf23 (lysB) are nonessential. Gene Y encodes a polypeptide with striking similarities to the family of holin proteins exemplified by gpS of phage lambda, and the Yam mutant displayed the expected properties of a holin mutant. The gene products of lysA and lysB, although nonessential, appear to play a role in the correct timing of lysis, since a lysA amber mutant caused slightly accelerated lysis and a lysB amber mutant slightly delayed lysis of nonpermissive strains. Gene X must encode a tail protein, since lysates from nonpermissive cells infected with the X amber mutant were complemented in vitro by similar lysates of cells infected with P2 head mutants but not with tail mutants.

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Complete Genome Sequence of Arthrobacter sp. ATCC 21022, a Host for Bacteriophage Discovery.

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