Molecular characterization of DNA puff II/9a genes in Sciara coprophila

DiBartolomeis, Susan M.; Gerbi, Susan A.

doi:10.1016/0022-2836(89)90129-0

Cited by 33 publications

(30 citation statements)

References 41 publications

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“…This paper describes a thorough analysis of a replication origin from Sciara DNA puff II/gA (locus 9A on chromosome II) by use of both the neutral/neutral and the neutral/alkaline 2D gel methods. Puff II/9A is the largest of the DNA puffs, contains two genes (II/9-1 and II/9-2) with 85% sequence similarity to each other that encode secretory polypeptides (DiBartolomeis and Gerbi 1989) and is amplified almost 20-fold (Wu et al 1993) This is the first time that 2D gels have been used successfully to map a replication origin in a multicellular eukaryote to a small region of no larger than 6 kb containing an -1-kb area where the majority of DNA replication initiates. Furthermore, replication is seen to proceed bidirectionally outward from this defined replication origin region.…”

mentioning

confidence: 99%

Replication initiates at a confined region during DNA amplification in Sciara DNA puff II/9A.

Liang¹,

Spitzer²,

Smith³

et al. 1993

Genes Dev.

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Two independent two-dimensional (2D) gel methods were used to map an origin of replication that is developmentally regulated by the steroid hormone ecdysone, namely an origin for DNA puff amplification in the fungus fly Sciara coprophila. Initiation of replication was found to occur within a small region of no larger than 6 kb by use of the neutral/neutral 2D gel method. Neutral/alkaline 2D gel analyses support the results of the neutral/neutral 2D gels and further define within the origin region an -l-kb area where the majority of replication initiates. This is the first example of an origin of replication in multicellular eukaryotes that has been mapped by 2D gels to such a small defined region. Moreover, replication can be seen by the neutral/alkaline 2D gel method to proceed bidirectionally outward from this replication origin region. These data are consistent with an onion-skin mechanism whereby multiple rounds of DNA replication initiate at a specific origin of replication for Sciara DNA puff amplification.

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mentioning

confidence: 99%

Replication initiates at a confined region during DNA amplification in Sciara DNA puff II/9A.

Liang¹,

Spitzer²,

Smith³

et al. 1993

Genes Dev.

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“…The Sciara DNA puff promoter was isolated as a 718-bp long EcoRV-Bal I fragment. It contains DNA sequences from -616 to +102 relative to the cap site of the gene (DiBartolomeis and Gerbi, 1989). This promoter fragment was ligated to the unique Sal I site (filled in by reverse transcriptase) of the polylinker of the P-element vector CarCAT-1 previously described by Mitsialis et al (1987).…”

Section: Materials and Methods Plasmid Construction And P-element Tramentioning

confidence: 99%

“…The 8.8-kb-long fragment of Sciara DNA puff 11/9A was isolated as a 1 .6-kb Xho I-Sal I fragment and a 7.2-kb Sal I-Sal I fragment (the downstream Sal I site comes from the X vector) (DiBartolomeis and Gerbi, 1989). The 1 .6-kbXho I-Sal I fragment was ligated into the unique Sal I site of the Carnegie 20 vector (Rubin and Spradling, 1983).…”

Section: Materials and Methods Plasmid Construction And P-element Tramentioning

confidence: 99%

“…The DNA binding site (EcRE) of the ecdysone receptor appears to be an imperfect palindrome, the more conserved right side of which has the consensus sequence of TGA A/C CY (Cherbas et al, 1991). There are three sequences (starting at -500, -421, and -271) (DiBartolomeis and Gerbi, 1989) in the 718-bp promoter fragment of DNA puff gene 11/9-1 identical to this putative EcRE consensus sequence except that Y is A/T in the Sciara sequences. This further agrees with the notion that the Sciara DNA puff promoter of gene 11/9-1 is directly controlled by the ecdysone receptor.…”

Section: C)mentioning

confidence: 99%

“…Unlike the cloned Drosophila genes from early-responding larval and prepupal RNA puffs (2B5, 74EF, and 75B) that encode regulatory proteins, DNA puff gene 11/9-1 seems to encode a structural protein. Isolation and characterization of DNA sequences derived from DNA puff locus 11/9A demonstrated the presence of two genes, 11/9-1 and 11/9-2, with 850/°sequence identity to each other (DiBartolomeis and Gerbi, 1989 …”

Section: Introductionmentioning

confidence: 99%

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The promoter of DNA puff gene II/9-1 of Sciara coprophila is inducible by ecdysone in late prepupal salivary glands of Drosophila melanogaster.

1991

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DNA puffs occur in Sciarid salivary gland chromosomes; they are sites of DNA amplification and intense transcription and they appear to encode secreted structural proteins needed for pupation. In this report we have used P-element transformation of Drosophila to study regulation of a Sciara DNA puff gene. We found that a 718-bp promoter fragment of DNA puff gene II/9-1 from Sciara coprophila directs expression of the bacterial reporter gene CAT in late prepupal salivary glands of transgenic Drosophila melanogaster. The identical tissue and analogous stage specificity indicate that some aspects of the ecdysone response are evolutionarily conserved between Drosophila and Sciara. When transgenic salivary glands are cultured in vitro, CAT activity is rapidly induced by ecdysone, suggesting direct control of gene expression by the ecdysone receptor. Putative stage-specific factors limit expression of the chimeric Sciara-CAT gene in transgenic Drosophila to late prepupae but not to third instar larvae when ecdysone titers are also high.

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The DNA puff 4C expresses a salivary secretion protein in Trichosia pubescens (Diptera; Sciaridae)

Andrioli

Gorab

Amabis

2007

Arch Insect Biochem Physiol

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DNA puffs are genomic regions of polytene chromosomes that undergo developmentally controlled DNA amplification and transcription in salivary glands of sciarid flies. Here, we tested the hypothesis that DNA puff genes code for salivary proteins in Trichosia pubescens. To do that, we generated antibodies against saliva and immunoscreened a cDNA library made from salivary glands. We isolated clones corresponding to DNA puff regions, including clone D-50 that contained the entire coding sequence of the previously isolated C4B1 gene from puff 4C. Indeed, we showed that puff 4C is a DNA puff region detecting its local transcription and its extra rounds of DNA incorporation compared to neighboring regions. We further confirmed D-50 clone identity in Western blots reacted with the anti-saliva anitiserum. We detected a recombinant protein expressed by this clone that had the expected size for a full-length product of the gene. We end with a discussion of the relationship between DNA puff genes and their products.

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Molecular characterization of DNA puff II/9a genes in Sciara coprophila

Cited by 33 publications

References 41 publications

Replication initiates at a confined region during DNA amplification in Sciara DNA puff II/9A.

Replication initiates at a confined region during DNA amplification in Sciara DNA puff II/9A.

The promoter of DNA puff gene II/9-1 of Sciara coprophila is inducible by ecdysone in late prepupal salivary glands of Drosophila melanogaster.

The DNA puff 4C expresses a salivary secretion protein in Trichosia pubescens (Diptera; Sciaridae)

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