The N-terminal domain of the insertion sequence 30 transposase interacts specifically with the terminal inverted repeats of the element

Rolf Stalder, Patrick Caspers, F. Olasz, Werner Arber

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The gene for the insertion sequence (IS)30 transposase is placed under the control of the tac promoter, and large quantities of transposase are expressed upon induction. The resulting protein precipitates inside the Escherichia coli cells in the form of inclusion bodies which, upon cell lysis, cannot be dissolved under non-denaturing conditions. In contrast, the N-terminal third of the transposase, a 17-kDa protein produced by a truncated gene, can be purified and is able to interact site specifically with the ends of the IS30 element. In DNase I footprint experiments, regions of 26 nucleotides on one DNA strand and 19 nucleotides on the other strand at either end of the element are protected from nuclease digestion. It is concluded that a functional DNA-binding domain can be formed by expression of only one-third of the complete IS30 transposase. Sequence comparison shows a homology of the IS30 ends to the ends of IS4351 and to the L1 end of bacteriophage Mu.

Original languageEnglish
Pages (from-to)3757-3762
Number of pages6
JournalJournal of Biological Chemistry
Volume265
Issue number7
Publication statusPublished - Mar 5 1990

Fingerprint

Transposases
Terminal Repeat Sequences
Insertional Mutagenesis
Nucleotides
Genes
Bacteriophages
Deoxyribonuclease I
DNA
Inclusion Bodies
Escherichia coli
Precipitates
Digestion
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

The N-terminal domain of the insertion sequence 30 transposase interacts specifically with the terminal inverted repeats of the element. / Stalder, Rolf; Caspers, Patrick; Olasz, F.; Arber, Werner.

In: Journal of Biological Chemistry, Vol. 265, No. 7, 05.03.1990, p. 3757-3762.

Research output: Contribution to journalArticle

@article{8b9e5c4166d840b6ad3a7d7b38baf678,
title = "The N-terminal domain of the insertion sequence 30 transposase interacts specifically with the terminal inverted repeats of the element",
abstract = "The gene for the insertion sequence (IS)30 transposase is placed under the control of the tac promoter, and large quantities of transposase are expressed upon induction. The resulting protein precipitates inside the Escherichia coli cells in the form of inclusion bodies which, upon cell lysis, cannot be dissolved under non-denaturing conditions. In contrast, the N-terminal third of the transposase, a 17-kDa protein produced by a truncated gene, can be purified and is able to interact site specifically with the ends of the IS30 element. In DNase I footprint experiments, regions of 26 nucleotides on one DNA strand and 19 nucleotides on the other strand at either end of the element are protected from nuclease digestion. It is concluded that a functional DNA-binding domain can be formed by expression of only one-third of the complete IS30 transposase. Sequence comparison shows a homology of the IS30 ends to the ends of IS4351 and to the L1 end of bacteriophage Mu.",
author = "Rolf Stalder and Patrick Caspers and F. Olasz and Werner Arber",
year = "1990",
month = "3",
day = "5",
language = "English",
volume = "265",
pages = "3757--3762",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "7",

}

TY - JOUR

T1 - The N-terminal domain of the insertion sequence 30 transposase interacts specifically with the terminal inverted repeats of the element

AU - Stalder, Rolf

AU - Caspers, Patrick

AU - Olasz, F.

AU - Arber, Werner

PY - 1990/3/5

Y1 - 1990/3/5

N2 - The gene for the insertion sequence (IS)30 transposase is placed under the control of the tac promoter, and large quantities of transposase are expressed upon induction. The resulting protein precipitates inside the Escherichia coli cells in the form of inclusion bodies which, upon cell lysis, cannot be dissolved under non-denaturing conditions. In contrast, the N-terminal third of the transposase, a 17-kDa protein produced by a truncated gene, can be purified and is able to interact site specifically with the ends of the IS30 element. In DNase I footprint experiments, regions of 26 nucleotides on one DNA strand and 19 nucleotides on the other strand at either end of the element are protected from nuclease digestion. It is concluded that a functional DNA-binding domain can be formed by expression of only one-third of the complete IS30 transposase. Sequence comparison shows a homology of the IS30 ends to the ends of IS4351 and to the L1 end of bacteriophage Mu.

AB - The gene for the insertion sequence (IS)30 transposase is placed under the control of the tac promoter, and large quantities of transposase are expressed upon induction. The resulting protein precipitates inside the Escherichia coli cells in the form of inclusion bodies which, upon cell lysis, cannot be dissolved under non-denaturing conditions. In contrast, the N-terminal third of the transposase, a 17-kDa protein produced by a truncated gene, can be purified and is able to interact site specifically with the ends of the IS30 element. In DNase I footprint experiments, regions of 26 nucleotides on one DNA strand and 19 nucleotides on the other strand at either end of the element are protected from nuclease digestion. It is concluded that a functional DNA-binding domain can be formed by expression of only one-third of the complete IS30 transposase. Sequence comparison shows a homology of the IS30 ends to the ends of IS4351 and to the L1 end of bacteriophage Mu.

UR - http://www.scopus.com/inward/record.url?scp=0025247877&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025247877&partnerID=8YFLogxK

M3 - Article

VL - 265

SP - 3757

EP - 3762

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 7

ER -