Faithful transcription initiation of fibroin gene in a homologous cell-free system reveals an enhancing effect of 5′ flanking sequence far upstream

Tsuda, Masashi; Suzuki, Yuzuru

doi:10.1016/0092-8674(81)90371-8

Cited by 128 publications

(86 citation statements)

References 11 publications

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“…However, such a modulating effect was not observed in the HeLa cell extract; the intensities of the 514-or 188-base bands were almost equal (compare lanes 1 and 2). These results are consistent with those obtained between the pFblOO and the 5'A-31 DNA described previously (12). Subsequently, we found that the intensity of the RNA bands from the pFblOO DNA and the 5'A-238 DNA was nearly indistinguishable (lanes 5 and 6), indicating that these two DNAs have an almost equal template activity and the region downstream from position -238 mainly accounts for the dominancy of the pFblOO DNA over the 5'A-44 DNA.…”

Section: Methodssupporting

confidence: 81%

“…1B). Specifically, in the silk gland extracts a modulating effect exerted by the sequences upstream from the TATA box has been observed (12). To detect the modulating effect on fibroin gene transcription, we have adopted a strategy to compare the template activities in one reaction mixture of two DNAs among several 5' deletion (A) genes, each of which has a different length of the 5' flanking sequence upstream from the TATA box (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Silk gland extracts were prepared from the posterior silk glands on the second day of the fifth larval instar by a procedure described previously (12). In vitro transcription assay and analysis of products also have been described (12 (15) and the silk gland extracts (12) requires a specific 35-bp stretch of DNA sequences (-29 to +6), including the TATA box (Fig. 1B).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, it is important to establish an in vitro transcription system that can reconstruct the effect of the DNA sequences upstream from the TATA box. We have recently found that the DNA sequences upstream from the TATA box modulate the template activity of the fibroin gene in a homologous cell-free system prepared from posterior silk glands (12), where the fibroin gene is actively transcribed (13).…”

mentioning

confidence: 99%

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Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box.

Tsuda¹,

Suzuki²

1983

Proc. Natl. Acad. Sci. U.S.A.

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We have previously reported that the 5'-flanking sequence upstream from the "TATA" box modulates the faithful transcription initiation of the fibroin gene in a homologous whole cell extract prepared from the silk glands, whereas such a modulating effect is not observed in a HeLa cell extract. Subsequently we have determined that major signals responsible for the modulating effect are located within a 200-base-pair region upstream from the TATA box, mainly in a distal region between nucleotide positions -238 and -116 and in a proximal one between -73 and -53. Inversion of the sequence element -234 to -66 did not alter its modulating effect. A similar modulating effect by the upstream region of the sericin gene was also observed in the silk gland extract but not in the HeLa cell extract. In contrast, a modulating effect by the upstream region of the adenovirus 2 major late gene was observed in the HeLa cell extract but not in the silk gland extract. Thus, the modulating effect by the upstream regions of these genes is exerted only in their own homologous extracts.

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Section: Methodssupporting

confidence: 81%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box.

Tsuda¹,

Suzuki²

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…We used HindIII-cleaved pU3RU5 as a truncated template which was expected to give rise to a run-off product of 330 bases upon the faithful initiation of transcription. To study specificity of transcription reactions catalyzed by various cell extracts, we also constructed plasmids containing three other well-characterized class II genes; the major late gene of Adenovirus 2 (Ad2ML) (19), the mouse 6-globin gene (20,21), and the mouse immunoglobulin VH101 gene (22) which should produce transcripts of about 560 bases, 470 bases, and 540 bases, respectively, when cleaved with restriction enzymes indicated in Fig. 1 Although these data support the preferential transcription of HTLV-I LTR in MT-2 extracts, the effect might be due to different -stabilities of RNA transcripts in various extracts.…”

Section: Construction Of Template Dnamentioning

confidence: 99%

Preferential transcription of HTLV-I LTR in cell-free extracts of human T cells producing HTLV-I viral proteins

Matsunami¹,

Siomi

Hatanaka

et al. 1986

Nucl Acids Res

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The promoters of the adenovirus 2 major late gene, the mouse S-globin gene, the mouse immunoglobulin VH gene and the LTR of the human T-lymphotropic retrovirus type I were tested for their transcription activities in cell-free extracts of four cell lines; HeLa, CESS (Epstein-Barr virustransformed human B cell line), MT-1 (HTLV-I-infected human T cell line without viral protein synthesis), and MT-2 (HTLV-I-infected human T cell line producing viral proteins). LTR was preferentially transcribed in the extracts of MT-2 although the other three genes were transcribed with relatively constant efficiencies in different extracts. The results agree well with the previous in vivo studies on the promoter activity of HTLV-I LTR. Mixing of HeLa and MT-2 extracts revealed the presence of a LTRspecific stimulating activity in MT-2 extracts.

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Sequences controlling in vitro transcription of SV40 promoters.

Hansen¹,

Sharp²

1983

The EMBO Journal

130

117

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A series of deletion mutants of SV40 were tested for early and late promoter activity in vitro in a transcription extract prepared from HeLa cells. These mutants had previously been characterized for expression in vivo. Transcription in vitro from both the SV40 early and late promoters was strongly dependent on an upstream region of DNA that contains six direct GC repeats. Sequences spanning two or more of these repeats stimulated transcription in a bidirectional fashion, at distances of 50‐200 bp. These sequences may function by mediating the activity of a specific transcriptional factor. Little effect on transcription in vitro was observed upon deletion of the 72‐bp enhancer elements. With this exception, the sequence dependence of early and late transcription in vitro was similar to that observed previously in vivo, both of the region including the GC repeats and of the early TATA sequence.

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Faithful transcription initiation of fibroin gene in a homologous cell-free system reveals an enhancing effect of 5′ flanking sequence far upstream

Cited by 128 publications

References 11 publications

Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box.

Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box.

Preferential transcription of HTLV-I LTR in cell-free extracts of human T cells producing HTLV-I viral proteins

Sequences controlling in vitro transcription of SV40 promoters.

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