Repetitive DNA sequences cotranscribed with developmentally regulated Dictyostelium discoideum mRNAs.

Zuker, Charles S.; Lodish, Harvey F.

doi:10.1073/pnas.78.9.5386

Cited by 63 publications

(31 citation statements)

References 28 publications

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“…In Further complications may arise from repeat sequences in different mRNAs. These have been shown to exist in slime molds (18,41) and possibly affect the selection procedures (22). Our selection procedures included higher criteri- Quantitative measurements of the steadystate concentrations showed that individual pluteus-specific mRNAs accumulated to different extents.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning of five individual stage- and tissue-specific mRNA sequences from sea urchin pluteus embryos.

Fregien¹,

Dolecki²,

Mandel³

et al. 1983

Mol. Cell. Biol.

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Five developmentally regulated sea urchin mRNA sequences which increase in abundance between the blastula and pluteus stages of development were isolated by molecular cloning of cDNA. The regulated sequences all appeared in moderately abundant mRNA molecules of pluteus cells and represented 4% of the clones tested. There were no regulated sequences detected in the 40% of the clones which hybridized to the most abundant mRNA, and the screening procedures were inadequate to detect possible regulation in the 20 to 30% of the clones presumably derived from rare-class mRNA. The reaction of 32P[cDNA] from blastula and pluteus mRNA to dots of the cloned DNAs on nitrocellulose filters indicated that the mRNAs complementary to the different cloned pluteus-specific sequences were between 3-and 47-fold more prevalent at the pluteus stage than at the blastula stage. Polyadenylated RNA from different developmental stages was transferred from electrophoretic gels to nitrocellulose filters and reacted to the different cloned sequences. The regulated mRNAs were undetectable in the RNA of 3-h embryos, became evident at the hatching blastula stage, and reached a maximum in abundance by the gastrula or pluteus stage. Certain of the clones reacted to two sizes of mRNA which did not vary coordinately with development. Transfers of RNA isolated from each of the three cell layers of pluteus embryos that were reacted to the cloned sequences revealed that two of the sequences were found in the mRNA of all three layers, two were ectoderm specific, and one was endoderm specific. Four of the regulated sequences were complementary to one or two major bands and one to at least 50 bands on Southern transfers of restriction endonuclease-digested total sea urchin DNA.The sea urchin embryo appears to be one of the most accessible developmental systems for studying the molecular events of cell type determination during early embryogenesis. After fertilization, eggs divide into a number of cells whose developmental fate is fixed somewhat before they differentiate into the various cell types. Most of the cells in the sea urchin embryo contribute to the three layers of the relatively simple pluteus larva: the ectodermal covering, the skeletal-forming mesenchyme, and the gut. These differentiated tissues function during the 1 to several months of larval life. The actual sea urchin emerges at metamorphosis after it has grown from a few cells set aside during embryogenesis for its development.Despite the simplicity of the system and the numerous molecular studies on sea urchin embryos, these larval tissues have not received much attention as specialized cells synthesizing cell type-specific proteins. The major reason has been that there is no evidence of very abundant With the advent of molecular cloning, it has become possible to define, isolate, and study genes coding for proteins produced only in limited abundance, including putative specialized proteins of the three cell layers of the pluteus. Clones of mRNA and genomic sequences which code for sev...

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Section: Discussionmentioning

confidence: 99%

Molecular cloning of five individual stage- and tissue-specific mRNA sequences from sea urchin pluteus embryos.

Fregien¹,

Dolecki²,

Mandel³

et al. 1983

Mol. Cell. Biol.

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“…Transcription of DIRS-1-related sequences results in the production of a heterogeneous population of polyadenylated cytoplasmic RNAs that are differentially expressed during development (7,24; C. Zuker, Ph.D. thesis, Massachusetts Institute of Technology, Cambridge). DIRS-1 RNAs are present at very low levels in growing cells, begin to accumulate within the first hour after initiation of development, and reach their maximal level by 15 h. Transcription of DIRS-1-related RNAs is induced in vegetative cells subjected to stresses such as heat shock and high cell density (22).…”

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confidence: 99%

“…Dictyostelium discoideum AX-3 was grown axenically and plated for development on filters as described previously (1). Growing cells were harvested when the concentration reached 2 x 106 cells per ml.The DIRS-1 genomic clones used here (pB41.6 and pB41.3) are subclones of the A genomic clone SB41 and have been described in detail elsewhere (7,22,24 …”

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confidence: 99%

“…The DIRS-1 genomic clones used here (pB41.6 and pB41.3) are subclones of the A genomic clone SB41 and have been described in detail elsewhere (7,22,24). pB41.3 contains the entire 4.1 kb of DIRS-1 unique internal sequence flanked by EcoRI sites.…”

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confidence: 99%

“…pB41.3 contains the entire 4.1 kb of DIRS-1 unique internal sequence flanked by EcoRI sites. pB41.6 contains a 2.5-kb EcoRI fragment that includes the right-terminal repeat of DIRS-1 and 2.2 kb of flanking sequence, which in SB41 is another copy of the right half of DIRS-1 (4,24).…”

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confidence: 99%

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Transcription of Dictyostelium discoideum transposable element DIRS-1.

Cohen¹,

Cappello²,

Lodish³

1984

Mol. Cell. Biol.

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DIRS-1 is a Dictyostelium discoideum transposable element that contains heat shock promoter sequences in the inverted terminal repeats. We showed that transcription of a 4.5-kilobase polyadenylated RNA initiates at a discrete site within the left-terminal repeat of DIRS-1, downstream from heat shock promoter and TATA box sequences. This RNA represents a full-length transcript of DIRS-1. We describe a cDNA clone that contains the 4.1 kilobases of internal sequence of DIRS-1, a cDNA clone that spans the junction between the internal sequences and the right-terminal repeat, and a cDNA clone that appears to have been transcribed from a rearranged genomic copy of DIRS-1. A second DIRS-1 RNA, named El, is transcribed on the opposite strand of DIRS-1 from the 4.5-kilobase RNA and is under control of the heat shock promoter in the right-terminal repeat. El transcription initiates at multiple positions both within and downstream from the right-terminal repeat. The same transcriptional initiation sites are used during normal development and during heat shock, suggesting that in all cases DIRS-1 transcription is regulated by the heat shock promoters contained within the two terminal repeats.Dictyostelium intermediate repeat sequence 1 (DIRS-1) is a moderately repetitive and apparently transposable element with several unusual features (4,7,15). DIRS-1 consists of 4.1 kilobases (kb) of unique internal sequence flanked by inverted terminal repeats of unequal length, 332 and 360 base pairs (bp) (6, 23). There are about 40 copies of the intact DIRS-1 element interspersed throughout the Dictyostelium genome, as well as an additional 200 copies of related sequences (7). Different Dictyostelium strains have different sequences flanking some of their DIRS-1 elements, indicating that the element had transposed after these strains were separated (7, 15).Transcription of DIRS-1-related sequences results in the production of a heterogeneous population of polyadenylated cytoplasmic RNAs that are differentially expressed during development (7, 24; C. Zuker, Ph.D. thesis, Massachusetts Institute of Technology, Cambridge). DIRS-1 RNAs are present at very low levels in growing cells, begin to accumulate within the first hour after initiation of development, and reach their maximal level by 15 h. Transcription of DIRS-1-related RNAs is induced in vegetative cells subjected to stresses such as heat shock and high cell density (22).Examination of the DNA sequence of the DIRS-1 terminal repeats has led to the identification of a putative heat shock promoter sequence (22, 23) homologous to the Drosophila consensus heat shock promoter (13,14). Stress-induced transcription of DIRS-1-related RNAs is presumed to be directed by these sequences. Cappello et al. (5) showed that a cloned partial copy of DIRS-1 (pB41.6) directs transcription of a heat-shock-inducible RNA in yeasts and that heatinducible transcription of this RNA is dependent on the presence of the DIRS-1 terminal repeat. The isolated terminal repeat also directs heat-inducible transcrip...

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Poly(A) RNA of the Egg Cytoplasm: Structural Resemblance to the Nuclear RNA of Somatic Cells

Davidson¹,

Jacobs²,

Thomas³

et al. 1983

Novartis Foundation Symposia

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Repetitive DNA sequences cotranscribed with developmentally regulated Dictyostelium discoideum mRNAs.

Cited by 63 publications

References 28 publications

Molecular cloning of five individual stage- and tissue-specific mRNA sequences from sea urchin pluteus embryos.

Molecular cloning of five individual stage- and tissue-specific mRNA sequences from sea urchin pluteus embryos.

Transcription of Dictyostelium discoideum transposable element DIRS-1.

Poly(A) RNA of the Egg Cytoplasm: Structural Resemblance to the Nuclear RNA of Somatic Cells

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