A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes

Singh, Harinder; Sen, Ranjan; Baltimore, David; Sharp, Phillip A.

doi:10.1038/319154a0

Cited by 1,146 publications

(584 citation statements)

References 30 publications

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“…Nuclear factor binding assays and polyacrylamide gel electrophoresis were performed essentially as described by Singh et al (14). Each assay contained 8 ,ug of nuclear extract protein (prepared by the method of Dignam et al [3]), 0.1 to 0.5 ng of the 32P-labeled DNA fragment to be analyzed, and 3.2 ,ug of poly(dIdC) poly(dI-dC) (Pharmacia).…”

Section: Methodsmentioning

confidence: 99%

Localization of transcriptional regulatory elements and nuclear factor binding sites in mouse ribosomal protein gene rpL32.

Atchison¹,

Meyuhas²,

Perry³

1989

Mol. Cell. Biol.

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The DNA sequences required for expression of the ribosomal protein gene rpL32 were identified by transient-expression assays of chimeric rpL32-chloramphenicol acetyltransferase genes. These studies showed that maximal rpL32 expression requires sequences in a 150-to 200-base-pair region spanning the transcriptional start site. Three discrete regions of importance were identified: one between positions -79 and -69 and two others located downstream of the transcriptional start site. Progressive 5' or 3' deletions caused stepwise decreases in expression, which suggested a complex interplay of redundant or compensatory elements. Gel mobility shift assays were used to idenfify trans-acting nuclear factors which bind to segments of the rpL32 promoter that are known to be important for transcription. Evidence for several distinct nuclear factors is presented. The binding sites for these factors were localized to the following regions: -79 to -69, -36 to -19, -19 to +11, +11 to +46 in exon I, and within the first 31 base pairs of intron 1. One of these factors may bind to multiple sites within the promoter region. Interestingly, the factor that binds to a sequence motif in the first exon also binds to similar motifs in a comparable region of the c-myc gene.The ribosomal proteins are structurally diverse and evolutionarily unrelated to each other, yet the genes that encode them are expressed with very similar efficiencies, as indicated by the relatively uniform abundance of various ribosomal protein mRNAs and by the similar RNA polymerase loading of different ribosomal protein genes (11, 12; D. E. Kelley and R. P. Perry, unpublished observations). What is the basis for this similarity? Presumably, the transcriptional efficiency of a ribosomal protein gene should ultimately be determined by the characteristics of its particular set of regulatory elements and by the cellular content of transacting factors that interact with these elements. To improve our understanding of these elements and factors, we have sought to define the DNA sequences that constitute the complete promoter of the ribosomal protein gene rpL32 and to identify the nuclear factors that bind to the functionally important sequences. Using a series of constructs in which selected portions of the rpL32 promoter region were linked to a chloramphenicol acetyltransferase (CAT) reporter gene, we observed that maximal expression requires sequences in a 150-to 200-base-pair (bp) region spanning the transcriptional start site, including sequences within exon I and intron 1. Moreover, we show that there are several discrete nuclear factor binding sites within this region, at least one of which recognizes a factor that is also bound by the myc gene promoter. These findings, together with previous and other parallel studies of rpL32 expression (2, 5; R. Moura-Neto, K. P. Dudov, and R. P. Perry, Proc. Natl. Acad. Sci. USA, in press), have delineated the complex set of sequences that determine the efficiency of rpL32 transcription. The unusual organization of this promote...

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Section: Methodsmentioning

confidence: 99%

Localization of transcriptional regulatory elements and nuclear factor binding sites in mouse ribosomal protein gene rpL32.

Atchison¹,

Meyuhas²,

Perry³

1989

Mol. Cell. Biol.

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“…In view of the unknown mechanism of enhancement, characterization of factors interacting with enhancer sequences is an important goal for the near future. Several reports about proteins interacting with enhancer sequences have been published recently (16,17,18,19,20,21). The enhancer of polyoma virus that is studied here is necessary both for transcription from the early promoter and for the replication of viral DNA (22,23,24).…”

Section: Intkroduionmentioning

confidence: 99%

Molecular analysis of the interaction between an enhancer binding factor and its DNA target

Piette

Yaniv

1986

Nucl Acids Res

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ABSTRATTIei fine contacts of a mouse nuclear factor, called PEB1, with the B enhancer of polyoma virus were analyzed. It protects against DNaseI attack a region of about 50 base pairs that can be divided in two domains. The first contains a GC-rich palindrome and the homology to the SV40 enhancer. The second is homologous to a sequence in the immunoglobulin (Ig) heavy chain gene enhancer. Methylation interference and protection experiments reveal strong specific contacts only with a purine rich track on the late coding strand of the early proximal part of the palindrome. Deletion analysis show that the minimal sequences necessary for binding include only the first domain. The Ig homology contributes only weakly to the binding. The minimal core is similar to the core of the B enhancer defined in vivo. The interactions we observe here are reminiscent of those of TFIIIA positive transcription factor and the 5SRNA gene of Xenopus. INTKRODUIONDNA-protein interactions play an essential role in the regulation of gene expression and DNA replication. Characterization of these interactions can help unravel the mechanisms involved in these processes. Repressor-operator interactions in prokaryotes were the first to be analyzed in great detail: in essence, a repressor dimer interacts with both parts of a palindromic sequence in a symmetric way; a conserved c-helix-turn-a-helix motif fits into the large groove of right-handed B-DNA (1). This model was found to be applicable to activator proteins like the CAP protein (2) or even yeast proteins involved in mating type control (3).In higher eukaryotes only a few cases of specific DNA-protein interactions have been studied in detail so far. The positive transcription factor TF III A of Xenopus laevis contacts the DNA mainly along the non coding strand (4) and the helical configuration of the DNA may be altered by this binding (5). Knowledge of the primary structure of the protein has led to a model in which small Zn++ binding, fingerlike domains interact with the DNA in a way quite different from bacterial repressors or activators (6). Another

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“…As a postdoctoral fellow with Phillip Sharp at the Massachussetts Institute of Technology, I was collaborating with members of David Baltimore's laboratory to discover and clone transcription factors that regulated the expression of Ig genes in B lymphocytes. During this period (1985)(1986)(1987)(1988)) I first extended the use of the electrophoretic mobility shift DNA binding assay, EMSA, to identify and characterize nuclear transcription factors (9). I then developed a novel method for cloning genes encoding such regulatory factors by screening lambda phage cDNA expression libraries with their DNA binding sites as probes (10).…”

Section: Pu1 a Shared Transcriptional Regulator Of Innate And Adaptmentioning

confidence: 99%

A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes

Cited by 1,146 publications

References 30 publications

Localization of transcriptional regulatory elements and nuclear factor binding sites in mouse ribosomal protein gene rpL32.

Localization of transcriptional regulatory elements and nuclear factor binding sites in mouse ribosomal protein gene rpL32.

Molecular analysis of the interaction between an enhancer binding factor and its DNA target

PU.1, a Shared Transcriptional Regulator of Innate and Adaptive Immune Cell Fates

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