Regulation of the replication of the murine immunoglobulin heavy chain gene locus: evaluation of the role of the 3' regulatory region

DNA replication in mammalian cells is a precisely controlled physical and temporal process, likely involving cis-acting elements that control the region(s) from which replication initiates. In B cells, previous studies showed replication timing to be early throughout the immunoglobulin heavy chain (Igh) locus. The implication from replication timing studies in the B-cell line MPC11 was that early replication of the Igh locus was regulated by sequences downstream of the C alpha gene. A potential candidate for these replication control sequences was the 3' regulatory region of the Igh locus. Our results demonstrate, however, that the Igh locus maintains early replication in a B-cell line in which the 3' regulatory region has been deleted from one allele, thus indicating that replication timing of the locus is independent of this region. In non-B cells (murine erythroleukemia cells [MEL]), previous studies of segments within the mouse Igh locus demonstrated that DNA replication likely initiated downstream of the Igh gene cluster. Here we use recently cloned DNA to demonstrate that segments located sequentially downstream of the Igh 3' regulatory region continue to replicate progressively earlier in S phase in MEL. Furthermore, analysis by two-dimensional gel electrophoresis indicates that replication forks proceed exclusively in the 3'-to-5' direction through the region 3' of the Igh locus. Extrapolation from these data predicts that initiation of DNA replication occurs in MEL at one or more sites within a 90-kb interval located between 40 and 130 kb downstream of the 3' regulatory region.

[1]  J. Diffley,et al.  ORC‐ and Cdc6‐dependent complexes at active and inactive chromosomal replication origins in Saccharomyces cerevisiae. , 1996, The EMBO journal.

[2]  J. Diffley,et al.  Once and only once upon a time: specifying and regulating origins of DNA replication in eukaryotic cells. , 1996, Genes & development.

[3]  M. Cogné,et al.  Palindromic structure of the IgH 3′locus control region , 1996, Nature Genetics.

[4]  D. Kioussis,et al.  Locus Control Region Function and Heterochromatin-Induced Position Effect Variegation , 1996, Science.

[5]  I. Simon,et al.  12 Temporal Order of DNA Replication , 1996 .

[6]  G. Wahl,et al.  Participation of the Human β-Globin Locus Control Region in Initiation of DNA Replication , 1995, Science.

[7]  E. Whitelaw,et al.  Position-dependent variegation of globin transgene expression in mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[8]  S. Bell.  Eukaryotic replicators and associated protein complexes. , 1995, Current opinion in genetics & development.

[9]  J. Michaelson,et al.  Identification of 3α-hs4, a novel Ig heavy chain enhancer element regulated at multiple stages of B cell differentiation , 1995 .

[10]  L. Madisen,et al.  Identification of a locus control region in the immunoglobulin heavy-chain locus that deregulates c-myc expression in plasmacytoma and Burkitt's lymphoma cells. , 1994, Genes & development.

[11]  F. Alt,et al.  A class switch control region at the 3′ end of the immunoglobulin heavy chain locus , 1994, Cell.

[12]  H. Cedar,et al.  Replication structure of the human & beta;-globin gene domain , 1993, Nature.

[13]  J. Yates,et al.  Constitutive binding of EBNA1 protein to the Epstein‐Barr virus replication origin, oriP, with distortion of DNA structure during latent infection. , 1993, The EMBO journal.

[14]  R. D. Little,et al.  Initiation and termination of DNA replication in human rRNA genes , 1993, Molecular and cellular biology.

[15]  K. Ariizumi,et al.  Immunoglobulin heavy chain enhancer is located near or in an initiation zone of chromosomal DNA replication. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Diffley,et al.  Protein-DNA interactions at a yeast replication origin , 1992, Nature.

[17]  P. Dijkwel,et al.  Mapping of replication initiation sites in mammalian genomes by two-dimensional gel analysis: stabilization and enrichment of replication intermediates by isolation on the nuclear matrix , 1991, Molecular and cellular biology.

[18]  M. Neuberger,et al.  The mouse IgH 3′‐enhancer , 1991, European journal of immunology.

[19]  L. Eckhardt,et al.  An enhancer at the 3' end of the mouse immunoglobulin heavy chain locus. , 1991, Nucleic acids research.

[20]  W. C. Forrester,et al.  A deletion of the human beta-globin locus activation region causes a major alteration in chromatin structure and replication across the entire beta-globin locus. , 1990, Genes & development.

[21]  M. Neuberger,et al.  A second B cell-specific enhancer 3' of the immunoglobulin heavy-chain locus , 1990, Nature.

[22]  C. Hutchison,et al.  Nucleotide sequence of the BALB/c mouse β-globin complex , 1989 .

[23]  M. Groudine,et al.  Asynchronous DNA replication within the human beta-globin gene locus. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[24]  C. Schildkraut,et al.  Replication program of active and inactive multigene families in mammalian cells , 1988, Molecular and cellular biology.

[25]  J. Huberman,et al.  Two-dimensional gel electrophoretic method for mapping DNA replicons , 1988, Molecular and cellular biology.

[26]  C. Schildkraut,et al.  Rate of replication of the murine immunoglobulin heavy-chain locus: evidence that the region is part of a single replicon , 1987, Molecular and cellular biology.

[27]  S. Morrison,et al.  Myeloma mutant with a novel 3' flanking region: loss of normal sequence and insertion of repetitive elements leads to decreased transcription but normal processing of the alpha heavy-chain gene products , 1986, Molecular and cellular biology.

[28]  N. Lehman,et al.  Purification of circular DNA using benzoylated naphthoylated DEAE-cellulose. , 1985, DNA.

[29]  C. Schildkraut,et al.  Changes in gene position are accompanied by a change in time of replication , 1984, Cell.