V(D)J recombinase binding and cleavage of cryptic recombination signal sequences identified from lymphoid malignancies

Ming Zhang, Patrick Swanson

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

V(D)J recombination is a process integral to lymphocyte development. However, this process is not always benign, since certain lymphoid malignancies exhibit recurrent chromosomal abnormalities, such as translocations and deletions, that harbor molecular signatures suggesting an origin from aberrant V(D)J recombination. Translocations involving LMO2, TAL1, Ttg-1, and Hox11, as well as a recurrent interstitial deletion at 1p32 involving SIL/SCL, are cited examples of illegitimate V(D)J recombination. Previous studies using extrachromosomal substrates reveal that cryptic recombination signal sequences (cRSSs) identified near the translocation breakpoint in these examples support V(D)J recombination with efficiencies ranging from about 30- to 20,000-fold less than bona fide V(D)J recombination signals. To understand the molecular basis for these large differences, we investigated the binding and cleavage of these cRSSs by the RAG1/2 proteins that initiate V(D)J recombination. We find that the RAG proteins comparably bind all cRSSs tested, albeit more poorly than a consensus RSS. We show that four cRSSs that support levels of V(D)J recombination above background levels in cell culture (LMO2, TAL1, Ttg-1, and SIL) are also cleaved by the RAG proteins in vitro with efficiencies ranging from 18 to 70% of a consensus RSS. Cleavage of LMO2 and Ttg-1 by the RAG proteins can also be detected in cell culture using ligation-mediated PCR. In contrast, Hox11 and SCL are nicked but not cleaved efficiently in vitro, and cleavage at other adventitious sites in plasmid substrates may also limit the ability to detect recombination activity at these cRSSs in cell culture.

Original languageEnglish
Pages (from-to)6717-6727
Number of pages11
JournalJournal of Biological Chemistry
Volume283
Issue number11
DOIs
StatePublished - Mar 14 2008

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VDJ Recombinases
V(D)J Recombination
Protein Sorting Signals
Genetic Recombination
Cell culture
RSS
Neoplasms
Cell Culture Techniques
Proteins
Lymphocytes
Substrates
Ports and harbors
Plasmids
Chromosome Aberrations
Ligation
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

V(D)J recombinase binding and cleavage of cryptic recombination signal sequences identified from lymphoid malignancies. / Zhang, Ming; Swanson, Patrick.

In: Journal of Biological Chemistry, Vol. 283, No. 11, 14.03.2008, p. 6717-6727.

Research output: Contribution to journalArticle

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