Functional organization of the inverted repeats of IS30

Mónika Szabó, János Kiss, F. Olasz

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The mobile element IS30 has 26-bp imperfect terminal inverted repeats (IRs) that are indispensable for transposition. We have analyzed the effects of IR mutations on both major transposition steps, the circle formation and integration of the abutted ends, characteristic for IS30. Several mutants show strikingly different phenotypes if the mutations are present at one or both ends and differentially influence the transposition steps. The two IRs are equivalent in the recombination reactions and contain several functional regions. We have determined that positions 20 to 26 are responsible for binding of the N-terminal domain of the transposase and the formation of a correct 2-bp spacer between the abutted ends. However, integration is efficient without this region, suggesting that a second binding site for the transposase may exist, possibly within the region from 4 to 11 bp. Several mutations at this part of the IRs, which are highly conserved in the IS30 family, considerably affected both major transposition steps. In addition, positions 16 and 17 seem to be responsible for distinguishing the IRs of related insertion sequences by providing specificity for the transposase to recognize its cognate ends. Finally, we show both in vivo and in vitro that position 3 has a determining role in the donor function of the ends, especially in DNA cleavage adjacent to the IRs. Taken together, the present work provides evidence for a more complex organization of the IS30 IRs than was previously suggested.

Original languageEnglish
Pages (from-to)3414-3423
Number of pages10
JournalJournal of Bacteriology
Volume192
Issue number13
DOIs
Publication statusPublished - Jul 2010

Fingerprint

Transposases
Mutation
DNA Cleavage
Terminal Repeat Sequences
Insertional Mutagenesis
Genetic Recombination
Binding Sites
Phenotype

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Functional organization of the inverted repeats of IS30. / Szabó, Mónika; Kiss, János; Olasz, F.

In: Journal of Bacteriology, Vol. 192, No. 13, 07.2010, p. 3414-3423.

Research output: Contribution to journalArticle

Szabó, Mónika ; Kiss, János ; Olasz, F. / Functional organization of the inverted repeats of IS30. In: Journal of Bacteriology. 2010 ; Vol. 192, No. 13. pp. 3414-3423.
@article{4fcb7adc837a48ed80e652c5611cb5fc,
title = "Functional organization of the inverted repeats of IS30",
abstract = "The mobile element IS30 has 26-bp imperfect terminal inverted repeats (IRs) that are indispensable for transposition. We have analyzed the effects of IR mutations on both major transposition steps, the circle formation and integration of the abutted ends, characteristic for IS30. Several mutants show strikingly different phenotypes if the mutations are present at one or both ends and differentially influence the transposition steps. The two IRs are equivalent in the recombination reactions and contain several functional regions. We have determined that positions 20 to 26 are responsible for binding of the N-terminal domain of the transposase and the formation of a correct 2-bp spacer between the abutted ends. However, integration is efficient without this region, suggesting that a second binding site for the transposase may exist, possibly within the region from 4 to 11 bp. Several mutations at this part of the IRs, which are highly conserved in the IS30 family, considerably affected both major transposition steps. In addition, positions 16 and 17 seem to be responsible for distinguishing the IRs of related insertion sequences by providing specificity for the transposase to recognize its cognate ends. Finally, we show both in vivo and in vitro that position 3 has a determining role in the donor function of the ends, especially in DNA cleavage adjacent to the IRs. Taken together, the present work provides evidence for a more complex organization of the IS30 IRs than was previously suggested.",
author = "M{\'o}nika Szab{\'o} and J{\'a}nos Kiss and F. Olasz",
year = "2010",
month = "7",
doi = "10.1128/JB.01382-09",
language = "English",
volume = "192",
pages = "3414--3423",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "13",

}

TY - JOUR

T1 - Functional organization of the inverted repeats of IS30

AU - Szabó, Mónika

AU - Kiss, János

AU - Olasz, F.

PY - 2010/7

Y1 - 2010/7

N2 - The mobile element IS30 has 26-bp imperfect terminal inverted repeats (IRs) that are indispensable for transposition. We have analyzed the effects of IR mutations on both major transposition steps, the circle formation and integration of the abutted ends, characteristic for IS30. Several mutants show strikingly different phenotypes if the mutations are present at one or both ends and differentially influence the transposition steps. The two IRs are equivalent in the recombination reactions and contain several functional regions. We have determined that positions 20 to 26 are responsible for binding of the N-terminal domain of the transposase and the formation of a correct 2-bp spacer between the abutted ends. However, integration is efficient without this region, suggesting that a second binding site for the transposase may exist, possibly within the region from 4 to 11 bp. Several mutations at this part of the IRs, which are highly conserved in the IS30 family, considerably affected both major transposition steps. In addition, positions 16 and 17 seem to be responsible for distinguishing the IRs of related insertion sequences by providing specificity for the transposase to recognize its cognate ends. Finally, we show both in vivo and in vitro that position 3 has a determining role in the donor function of the ends, especially in DNA cleavage adjacent to the IRs. Taken together, the present work provides evidence for a more complex organization of the IS30 IRs than was previously suggested.

AB - The mobile element IS30 has 26-bp imperfect terminal inverted repeats (IRs) that are indispensable for transposition. We have analyzed the effects of IR mutations on both major transposition steps, the circle formation and integration of the abutted ends, characteristic for IS30. Several mutants show strikingly different phenotypes if the mutations are present at one or both ends and differentially influence the transposition steps. The two IRs are equivalent in the recombination reactions and contain several functional regions. We have determined that positions 20 to 26 are responsible for binding of the N-terminal domain of the transposase and the formation of a correct 2-bp spacer between the abutted ends. However, integration is efficient without this region, suggesting that a second binding site for the transposase may exist, possibly within the region from 4 to 11 bp. Several mutations at this part of the IRs, which are highly conserved in the IS30 family, considerably affected both major transposition steps. In addition, positions 16 and 17 seem to be responsible for distinguishing the IRs of related insertion sequences by providing specificity for the transposase to recognize its cognate ends. Finally, we show both in vivo and in vitro that position 3 has a determining role in the donor function of the ends, especially in DNA cleavage adjacent to the IRs. Taken together, the present work provides evidence for a more complex organization of the IS30 IRs than was previously suggested.

UR - http://www.scopus.com/inward/record.url?scp=77954366519&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77954366519&partnerID=8YFLogxK

U2 - 10.1128/JB.01382-09

DO - 10.1128/JB.01382-09

M3 - Article

C2 - 20418401

AN - SCOPUS:77954366519

VL - 192

SP - 3414

EP - 3423

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 13

ER -