Distinct DNA sequence and structure requirements for the two steps of V(D)J recombination signal cleavage

Dale A. Ramsden; J. Fraser McBlane; Dik C. Van Gent; Martin Geliert

doi:10.1002/j.1460-2075.1996.tb00682.x

Distinct DNA sequence and structure requirements for the two steps of V(D)J recombination signal cleavage

Dale A. Ramsden, J. Fraser McBlane, Dik C. Van Gent, Martin Geliert^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

157 Citations (Scopus)

Abstract

Cleavage of V(D)J recombination signals by purified RAG1 and RAG2 proteins permits the dissection of DNA structure and sequence requirements. The two recognition elements of a signal (nonamer and heptamer) are used differently, and their cooperation depends on correct helical phasing. The nonamer is most important for initial binding, while efficient nicking and hairpin formation require the heptamer sequence. Both nicking and hairpin formation are remarkably tolerant of variations in DNA structure. Certain flanking sequences inhibit hairpin formation, but this can be bypassed by base unpairing, and even a completely single-stranded signal sequence is well utilized. We suggest that DNA unpairing around the signal-coding border is essential for the initiation of V(D)J recombination.

Original language	English
Pages (from-to)	3197-3206
Number of pages	10
Journal	EMBO Journal
Volume	15
Issue number	12
DOIs	https://doi.org/10.1002/j.1460-2075.1996.tb00682.x
Publication status	Published - 17 Jun 1996
Externally published	Yes

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10.1002/j.1460-2075.1996.tb00682.x

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title = "Distinct DNA sequence and structure requirements for the two steps of V(D)J recombination signal cleavage",

abstract = "Cleavage of V(D)J recombination signals by purified RAG1 and RAG2 proteins permits the dissection of DNA structure and sequence requirements. The two recognition elements of a signal (nonamer and heptamer) are used differently, and their cooperation depends on correct helical phasing. The nonamer is most important for initial binding, while efficient nicking and hairpin formation require the heptamer sequence. Both nicking and hairpin formation are remarkably tolerant of variations in DNA structure. Certain flanking sequences inhibit hairpin formation, but this can be bypassed by base unpairing, and even a completely single-stranded signal sequence is well utilized. We suggest that DNA unpairing around the signal-coding border is essential for the initiation of V(D)J recombination.",

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T1 - Distinct DNA sequence and structure requirements for the two steps of V(D)J recombination signal cleavage

AU - Ramsden, Dale A.

AU - McBlane, J. Fraser

AU - Van Gent, Dik C.

AU - Geliert, Martin

PY - 1996/6/17

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AB - Cleavage of V(D)J recombination signals by purified RAG1 and RAG2 proteins permits the dissection of DNA structure and sequence requirements. The two recognition elements of a signal (nonamer and heptamer) are used differently, and their cooperation depends on correct helical phasing. The nonamer is most important for initial binding, while efficient nicking and hairpin formation require the heptamer sequence. Both nicking and hairpin formation are remarkably tolerant of variations in DNA structure. Certain flanking sequences inhibit hairpin formation, but this can be bypassed by base unpairing, and even a completely single-stranded signal sequence is well utilized. We suggest that DNA unpairing around the signal-coding border is essential for the initiation of V(D)J recombination.

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Distinct DNA sequence and structure requirements for the two steps of V(D)J recombination signal cleavage

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