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Titolo:
STIMULATION OF DEFECTIVE-DNA TRANSFER ACTIVITY IN RECOMBINATION DEFICIENT SCID CELL-EXTRACTS BY A 72-KDA PROTEIN FROM WILD-TYPE THYMOCYTES
Autore:
JESSBERGER R; RIWAR B; ROLINK A; RODEWALD HR;
Indirizzi:
BASEL INST IMMUNOL,GRENZACHERSTR 487 CH-4005 BASEL SWITZERLAND
Titolo Testata:
The Journal of biological chemistry
fascicolo: 12, volume: 270, anno: 1995,
pagine: 6788 - 6797
SICI:
0021-9258(1995)270:12<6788:SODTAI>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
STRAND-BREAK REPAIR; LYMPHOID V(D)J RECOMBINATION; MOUSE L-CELLS; GENE REARRANGEMENT; MOLECULAR-CLONING; ANTIGEN RECEPTOR; SIGNAL SEQUENCE; CODING SEGMENTS; KINASE; MECHANISM;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
62
Recensione:
Indirizzi per estratti:
Citazione:
R. Jessberger et al., "STIMULATION OF DEFECTIVE-DNA TRANSFER ACTIVITY IN RECOMBINATION DEFICIENT SCID CELL-EXTRACTS BY A 72-KDA PROTEIN FROM WILD-TYPE THYMOCYTES", The Journal of biological chemistry, 270(12), 1995, pp. 6788-6797

Abstract

The SCID (Severe Combined Immune Deficiency) mutation causes two DNA recombination deficiencies: an aberrant joining of V(D)J immunoglobulin gene elements and a failure to perform efficient repair of DNA double-strand breaks. A recently established cell-free assay for DNA transfer (DTA) was applied to study nuclear extracts from normal and SCID-derived cells. The recombination deficiency was reflected in the cell-free system: SCID lymphocyte and fibroblast extracts showed reduced levels of DTA activity on a variety of DNA substrates. Analysis of nuclearextracts prepared from wild-type thymocytes and B cells representing different stages in lymphocyte ontogeny revealed the highest activities at the most immature stages. With progression of development, DTA activity decreased. Corresponding to their early developmental arrest, V(D)J rearrangement-incompetent RAG-2(-/-) lymphocyte extracts show high DTA activity. In contrast, extracts from SCID early lymphocytes express very low DNA transfer activity. Induction of V(D)J rearrangement in vivo in a normal preB cell line lead to a co-induction of the cell-free recombination activity. This indicates a development stage specificity of cell-free DNA recombination, which temporally parallels V(D)J recombination. A protein could be purified to near-homogeneity from wild-type thymocytes which stimulates the recombination activity specifically in SCID thymocyte and proB cell extracts. This protein, SRSP (SCID Recombination Stimulatory Protein), migrates as a single band of approximately 72 kDa in SDS-polyacrylamide gel electrophoresis.

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Documento generato il 27/11/20 alle ore 10:10:40