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      The zinc-finger protein Zelda is a key activator of the early zygotic genome in Drosophila

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          Abstract

          In all animals, the initial events of embryogenesis are controlled by maternal gene products that are deposited into the developing oocyte. At some point after fertilization, control of embryogenesis is transferred to the zygotic genome in a process called the maternal to zygotic transition (MZT). During this time many maternal RNAs are degraded and transcription of zygotic RNAs ensues 1. A longstanding question has been, what factors regulate these events? The recent findings that microRNAs 2, 3 and Smaugs 4 mediate maternal transcript degradation have shed new light on this aspect of the problem. However, the transcription factor(s) that activate the zygotic genome remain elusive. The discovery that many of the early transcribed genes in Drosophila share a cis-regulatory heptamer motif, CAGGTAG and related sequences 5, 6, collectively referred to as TAGteam sites 5 brought up the possibility that a dedicated transcription factor could interact with these sites to activate transcription. Here we report that the zinc-finger protein, Zelda (Zld; Zinc-finger ear ly Drosophila activator), binds specifically to these sites, and is capable of activating transcription in transient transfection assays. Mutant embryos lacking zld are defective in cellular blastoderm formation, and fail to activate many genes essential for cellularization, sex determination, and pattern formation. Global expression profiling confirmed that Zld plays a key role in the activation of the early zygotic genome, and suggests that Zld may also regulate maternal RNA degradation during the MZT.

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          Most cited references25

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          The DrosDel collection: a set of P-element insertions for generating custom chromosomal aberrations in Drosophila melanogaster.

          We describe a collection of P-element insertions that have considerable utility for generating custom chromosomal aberrations in Drosophila melanogaster. We have mobilized a pair of engineered P elements, p[RS3] and p[RS5], to collect 3243 lines unambiguously mapped to the Drosophila genome sequence. The collection contains, on average, an element every 35 kb. We demonstrate the utility of the collection for generating custom chromosomal deletions that have their end points mapped, with base-pair resolution, to the genome sequence. The collection was generated in an isogenic strain, thus affording a uniform background for screens where sensitivity to genetic background is high. The entire collection, along with a computational and genetic toolbox for designing and generating custom deletions, is publicly available. Using the collection it is theoretically possible to generate >12,000 deletions between 1 bp and 1 Mb in size by simple eye color selection. In addition, a further 37,000 deletions, selectable by molecular screening, may be generated. We are now using the collection to generate a second-generation deficiency kit that is precisely mapped to the genome sequence.
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            A major developmental transition in early Xenopus embryos: II. Control of the onset of transcription.

            We have shown in the accompanying paper that a developmental transition occurs at the midblastula stage (cleavage 12) in Xenopus embryos, and that this midblastula transition (MBT) is apparently initiated when the ratio of nucleus to cytoplasm reaches a critical value. One manifestation of this transition is the onset of transcription. We show here that a plasmid containing a cloned gene coding for a yeast leucine tRNA comes under developmental control when injected into cleaving eggs. In pre-MBT eggs this plasmid is transiently transcribed and then becomes inactive; however, it becomes transcriptionally active again at the MBT. This pre-MBT suppression of transcription can be reversed by addition of competing DNA. The amount of DNA needed to induce premature transcription is equal to the amount of nuclear DNA present after 12 cleavages (24 ng), suggesting that the MBT is triggered by the DNA through titration of suppressor components present in the egg.
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              The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster.

              The production of female germline chimeras is invaluable for analyzing the tissue specificity of recessive female sterile mutations as well as detecting the maternal effect of recessive zygotic lethal mutations. Previously, we developed the "FLP-DFS" technique to efficiently generate germline clones. This technique uses the X-linked germline-dependent dominant female sterile mutation ovoD1 as a selection for the detection of germline recombination events, and the FLP-FRT recombination system to promote site-specific chromosomal exchange. This method allows the efficient production of germline mosaics only on the X chromosome. In this paper we have built chromosomes that allow the use of this technique to the autosomes. We describe the various steps involved in the development of this technique as well as the properties of the chromosomes utilized.
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                Author and article information

                Journal
                0410462
                6011
                Nature
                Nature
                0028-0836
                1476-4687
                4 September 2008
                19 October 2008
                20 November 2008
                20 May 2009
                : 456
                : 7220
                : 400-403
                Affiliations
                [1 ] Department of Biology, New York University, 100 Washington Square East, New York, NY 10003, USA.
                [2 ] Institute of Human Genetics, University of Utah, Salt Lake City, UT 84112, USA.
                Author notes
                [*]

                These authors contributed equally to this work

                Author Information The microarray data discussed in this publication have been deposited in NCBI’s Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) and are accessible through GEO series accession number GSE11231. Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests. Correspondence and requests for materials should be addressed to C. R. ( chris.rushlow@ 123456nyu.edu ).

                Article
                nihpa67900
                10.1038/nature07388
                2597674
                18931655
                26aaa56d-ecd9-46ed-8aec-81b1e5a2b0fa
                History
                Funding
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM063024-05 ||GM
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM063024-04 ||GM
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM063024-03 ||GM
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM063024-02 ||GM
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM063024-01A1 ||GM
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