The primary transcription unit of the mouse β-Major globin gene

Hofer, Erhard; Darnell, James

doi:10.1016/0092-8674(81)90154-9

Cited by 331 publications

(147 citation statements)

References 33 publications

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“…The same Ul gene is initiated at base 1 wherl it is injected into Xenopus oocytes (35). In addition, there is a minor transcription start site at -110 in the Xenopus oocyte which yields a stable RNA which can be precipitated with anti-RNP antibody (35) (16,25). Nonpolyadenylated RNAs such as histone mRNAs also are formed by cleavage of the transcript at the 3' end (1,32).…”

Section: Discussionmentioning

confidence: 99%

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Morris¹,

Marzluff²

1985

Mol. Cell. Biol.

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Nuclei from sea urchin blastula embryos synthesize a variety of small RNAs, one of which has identical mobility with sea urchin Ul RNA. This RNA is synthesized by RNA polymerase II and, in a hybridizationselection experiment, was selected by the cloned sea urchin Ul gene. The Ul RNA was initiated with ATP, but not GIP, in isolated nuclei with P-S-and y-S-ribonucleotide triphosphates as substrates. The Ul RNA containing thiophosphate at the 5' end was not capped but accumulated as an uncapped transcript from which the thiophosphate could be removed with calf intestinal phosphatase.

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Section: Discussionmentioning

confidence: 99%

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Morris¹,

Marzluff²

1985

Mol. Cell. Biol.

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“…Several eukaryotic genes have been shown to be transcribed beyond their polyadenylylation site (20)(21)(22)(23)(24)(25)(26). If this is also the case for the growth hormone gene, the primary growth hormone transcript would have to be processed, possibly by a specialized endonuclease, before the poly(A) tail can be added onto the molecule.…”

Section: Discussionmentioning

confidence: 99%

Requirement for the 3' flanking region of the bovine growth hormone gene for accurate polyadenylylation.

Woychik¹,

Lyons²,

Post³

et al. 1984

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

110

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We examined whether the sequence extending 3' to the polyadenylylation site of the bovine growth hormone gene contains any signal that affects the polyadenylylation of the growth hormone mRNA. For this purpose, cloned copies of this gene, each containing a different length of growth hormone-specific sequence 3' to the wild-type polyadenylylation site, were used to transfect COS-1 cells. The polyadenylylation site on the mRNAs produced from the exogenously added growth hormone genes were analyzed with an S1 nuclease mapping procedure. We found that a gene containing 84 base pairs of its own 3' flanking sequence is capable of producing an accurately polyadenylylated mRNA. On the other hand, genes containing only 1, 10, or 13 base pairs of 3' flanking sequence were principally polyadenylylated at discrete sites either upstream or downstream from the wild-type position. Using a computer program, we examined whether secondary structures on the primary growth hormone transcript correlated with the site where the mRNA is polyadenylylated.

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“…CAT activity was measured by thin-layer chromatography (19), and f8-galactosidase activity was assayed in an ortho-nitrophenyl f3-D-galactopyranoside (Sigma) as substrate (20). RNA chain elongation in isolated nuclei was performed as described (21).…”

Section: Methodsmentioning

confidence: 99%

Transcriptional control of the endogenous MYC protooncogene by antisense RNA.

Yokoyama

Imamoto

1987

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

129

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DNAs (20 ,ug) by the protoplast fusion method (10). Two days later, selection for the ability to grow in Dulbecco's modified Eagle's medium (DMEM) containing mycophenolic acid (25 ,gg/ml), aminopterin, xanthine, hypoxanthine, and thymidine was performed as described by Mulligan and Berg (11). Transformants resistant to a high dose of mycophenolic acid (110 Ag/ml) were obtained by culturing cells in DMEM plus mycophenolic acid over a period of 6 months during which the mycophenolic acid concentration was increased at 2-week intervals. Secondary and tertiary transformants were obtained by retransfecting a primary and a secondary transformant, respectively, with plasmid DNA (10 ,ug/ml) followed by culturing in DMEM containing mycophenolic acid (110 ,g/ml). Control antisense and sense clones (pSV2gpt+, pSVgptC5-8, pSVH-2Kb, and pSVa-globin) were obtained by the cotransfection with pSV2gpt+. The same amplification protocol and the repeated transfection were used as described above.RNA Isolation and Analysis. Total RNA, poly(A)+ mRNA, and cytoplasmic and nuclear RNAs were prepared by the standard protocols (12, 13). RNA gel blot transfer and hybridization were carried out as described by Maniatis et al.(13). [a-32P]dCTP-labeled single-stranded probes were prepared from restriction fragments containing f,-actin cDNA (14) or exons 2 and 3 of MYC cDNA by exonuclease III digestion, reverse transcriptase treatment (15), and strand separation (13 7363The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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The primary transcription unit of the mouse β-Major globin gene

Cited by 331 publications

References 33 publications

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Requirement for the 3' flanking region of the bovine growth hormone gene for accurate polyadenylylation.

Transcriptional control of the endogenous MYC protooncogene by antisense RNA.

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