Regulation of Mu transposition II. The Escherichia coli HimD protein positively controls two repressor promoters and the early promoter of bacteriophage Mu

Goosen, Nora; Heuvel, Margreet van; Moolenaar, Geri F.; Putte, Pieter van de

doi:10.1016/0378-1119(84)90017-9

Cited by 59 publications

(45 citation statements)

References 13 publications

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“…With 8-min RNA, one hybridization signal predominated (Fig. 2), indicating transcription initiation within fragment 7, which contains the early promoter Pe and the repressor promoter PcM (11,22,23 (27) except that the sizes for fragments 46 (0.97 kb) and 48 Effects of mutations on Mu promoter activity. The three phage mutations AamlO93, BamlO66, and A130 (a Mu rightend deletion removing attR), which prevent Mu-specific DNA replication (6, 50), had no discernible effect on early transcript initiation (Fig.…”

Section: Methodsmentioning

confidence: 98%

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Localization and regulation of bacteriophage Mu promoters

Stoddard

Howe

1989

J Bacteriol

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on Southern blots. Signals were detected from four new promoters in addition to the previously characterized Pe (early), PCM (repressor), and P,om (late) promoters. A major signal upstream of C was first observed at 12 min and intensified thereafter with RNA from cts and C amber but not replication-defective prophages; these characteristics indicate that this signal arises from a middle promoter, which we designate Pm. 40-min RNA, four additional major signals originated in the C-lys, F-G-I, N-P, and com-mom regions. These signals were missing with RNA from C amber and replication-defective prophages and therefore reflected the activity of late promoters, one of which we presume was Po,,, Uninduced lysogens showed weak signals from five regions, one from the early regulatory region, three between genes B and lys, and one near the late genes K, L, and M. The first of these probably resulted from PCM activity; the others remain to be identified.

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Section: Methodsmentioning

confidence: 98%

“…scribed leftward from the PcM promoter, located -1 kilobase (kb) from the left end (10,11,22). Studies with in vivo pulse-labeled RNAs isolated after heat induction of Mu cts prophages have revealed three transcriptional phases during lytic development: early, middle, and late (10; C. F. Marrs and M. M. Howe, submitted for publication).…”

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confidence: 99%

Localization and regulation of bacteriophage Mu promoters

Stoddard

Howe

1989

J Bacteriol

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“…That a small protein like Ner protects such a large region is in sharp contrast to larger proteins such as A and 434 repressors [20,21], whose interactions span smaller regions of DNA. Mu ner operator region II, encompasses the transcriptional initiation site of the repressor promoter PCZ [22], whereas II, spans a turn of the DNA that includes the ' -10' promoter region of the early promoter Pe [7]. Upon binding, Ner may ultimately mediate its action by sterically hindering the initiation of transcription from both PCZ and Pe.…”

Section: Discussionmentioning

confidence: 99%

“…Integration host factor (IHF), comprised of the himA and himD gene products [23], is a heterodimer which positively regulates expression from both PCZ and Pe [22,24]. Expression from wild type Pe is markedly reduced in the absence of IHF [22].…”

Section: Discussionmentioning

confidence: 99%

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Purification and characterization of the Ner repressor of bacteriophage Mu

Kukolj¹,

Tolias²,

DuBow³

1989

FEBS Letters

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The Ner protein of bacteriophage Mu acts as a I cro-like negative regulator of the phage's early (transposase) operon. Using the band retardation assay to monitor ner-operator-specific DNA-binding activity, the 8 kDa Ner protein was purified to homogeneity. DNase I footprinting revealed that the purified protein bound and protected a specific DNA operator that contains two 12 bp sites with the consensus sequence S-ANPyTAPuCTAAGT-3', separated by a 6 bp spacer region. Moreover, regions corresponding to a turn of the DNA helix flanking these 12 bp repeats are also protected by Ner. Unlike the functionally similar I cro protein, gel filtration experiments show the native molecular mass of Mu Ner to be approx. 8 kDa. These results, plus the pattern of DNase I protection, suggest that the protein may bind as a monomer to each of its specific DNA substrates.Bacteriophage Mu; Protein, Ner; DNA-protein interaction

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Escherichia coli integration host factor bends the DNA at the ends of IS1 and in an insertion hotspot with multiple IHF binding sites.

1987

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The integration host factor of Escherichia coli (IHF) is a small, histone‐like protein which participates in the integration of bacteriophage lambda into the E. coli chromosome and in a number of regulatory processes. Our recent footprinting analysis has shown that IHF binds specifically to the ends of the transposable element IS1, as well as to several sites within a short segment of the plasmid pBR322. We have extended our studies of the binding of the IHF molecule to these sites in vitro using a gel retardation assay. We report here that IHF bends the DNA upon binding, as judged from the strong cyclic dependence of the protein‐induced mobility shift on the position of the binding site. Using cloned, synthetic ends of IS1 as substrates, we have found that some mutations within the conserved bases of the IHF consensus binding sequence abolish binding, and that alterations of the flanking sequences can greatly reduce IHF binding. The presence of multiple IHF sites on a single DNA fragment increases binding very little, indicating that IHF does not bind cooperatively in this complex. We discuss the possibility that DNA bending is related to the role IHF plays in forming and stabilizing nucleoprotein complexes, and suggest that bending at the IHF sites may be important to its diverse effects in the cell.

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Regulation of Mu transposition II. The Escherichia coli HimD protein positively controls two repressor promoters and the early promoter of bacteriophage Mu

Cited by 59 publications

References 13 publications

Localization and regulation of bacteriophage Mu promoters

Localization and regulation of bacteriophage Mu promoters

Purification and characterization of the Ner repressor of bacteriophage Mu

Escherichia coli integration host factor bends the DNA at the ends of IS1 and in an insertion hotspot with multiple IHF binding sites.

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