New baculovirus expression tools for recombinant protein complex production

Trowitzsch, Simon; Bieniossek, Christoph; Nie, Yan; Garzoni, Frédéric; Berger, Imre

doi:10.1016/j.jsb.2010.02.010

Cited by 186 publications

(191 citation statements)

References 69 publications

(99 reference statements)

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“…The 5Ј region of the GCN5 gene was engineered to code for a decahistidine tag followed by a tobacco etch virus protease cleavage site. Vectors were fused by Cre recombination and integrated into the EMBacY baculovirus genome via Tn7 transposition (44). Recombinant baculoviruses were generated as described (44) and used for protein complex production in Sf21 insect cell culture.…”

Section: Dna Constructs and Recombinant Protein Production-thementioning

confidence: 99%

“…Vectors were fused by Cre recombination and integrated into the EMBacY baculovirus genome via Tn7 transposition (44). Recombinant baculoviruses were generated as described (44) and used for protein complex production in Sf21 insect cell culture. Infected insect cells were harvested 72 h post cell arrest by centrifugation and stored at Ϫ80°C until further use.…”

Section: Dna Constructs and Recombinant Protein Production-thementioning

confidence: 99%

“…Subunits corresponding to either the HAT module of hATAC (GCN5, ADA2a, ADA3, and SGF29) or those of the HAT modules of hSAGA (GCN5, ADA2b, ADA3, and SGF29) were produced in insect cells using the MultiBac expression system (44,49). Recombinant proteins were purified from extracts of MultiBac-infected Sf21 insect cells by double immunoprecipitation against hGCN5 and ADA3 (Fig.…”

Section: Purification Of Gcn5-and Gcn5-containing Complexes-mentioning

confidence: 99%

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Subunits of ADA-two-A-containing (ATAC) or Spt-Ada-Gcn5-acetyltrasferase (SAGA) Coactivator Complexes Enhance the Acetyltransferase Activity of GCN5

Riss

Scheer

Joint

et al. 2015

Journal of Biological Chemistry

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Background: Histone acetyltransferases (HATs) incorporated in large multiprotein complexes are involved in a wide variety of cellular processes, including transcription regulation. Results: Subunits of HAT complexes enhance the enzymatic activity of their catalytic subunits. Conclusion: The activity, but not the specificity of HAT complexes, is stimulated by the corresponding subunits. Significance: We gained important insights into histone acetylation function and specificity of HAT complexes.

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Section: Dna Constructs and Recombinant Protein Production-thementioning

confidence: 99%

Section: Dna Constructs and Recombinant Protein Production-thementioning

confidence: 99%

Section: Purification Of Gcn5-and Gcn5-containing Complexes-mentioning

confidence: 99%

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Subunits of ADA-two-A-containing (ATAC) or Spt-Ada-Gcn5-acetyltrasferase (SAGA) Coactivator Complexes Enhance the Acetyltransferase Activity of GCN5

Riss

Scheer

Joint

et al. 2015

Journal of Biological Chemistry

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“…Within the SPINE2-COMPLEXES consortium a wide panel of cloning strategies and vector sets have been developed to streamline construct design for expression/co-expression screening in Escherichia coli (Busso et al, 2005;de Jong et al, 2006;Berrow et al, 2007;Scheich et al, 2007;Fogg and Wilkinson, 2008;Bieniossek et al, 2009;Unger et al, 2010;Diebold et al, 2011) (Luna-Vargas et al, 2011 as well as in insect and mammalian cells (Aricescu et al, 2006;Berrow et al, 2007;Abdulrahman et al, 2009;Pradeau-Aubreton et al, 2010;Trowitzsch et al, 2010). A variety of new technologies for DNA manipulation including ligation independent or restriction free procedures, in-fusion or gateway approaches are now being used in addition to classical restriction-based strategies (see Busso et al, 2011 for examples and test cases).…”

Section: Challenges For Sample Preparation: New Methods For Protein Cmentioning

confidence: 99%

Structural insights into transcription complexes

Berger

Blanco

Boelens

et al. 2011

Journal of Structural Biology

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a b s t r a c tControl of transcription allows the regulation of cell activity in response to external stimuli and research in the field has greatly benefited from efforts in structural biology. In this review, based on specific examples from the European SPINE2-COMPLEXES initiative, we illustrate the impact of structural proteomics on our understanding of the molecular basis of gene expression. While most atomic structures were obtained by X-ray crystallography, the impact of solution NMR and cryo-electron microscopy is far from being negligible. Here, we summarize some highlights and illustrate the importance of specific technologies on the structural biology of protein-protein or protein/DNA transcription complexes: structure/ function analysis of components the eukaryotic basal and activated transcription machinery with focus on the TFIID and TFIIH multi-subunit complexes as well as transcription regulators such as members of the nuclear hormone receptor families. We also discuss molecular aspects of promoter recognition and epigenetic control of gene expression.

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“…The rising of protein-protein interaction studies as well as the constantly increasing demand of proteins in drug discovery programs have brought the need for high-throughput protein expression and automation to the foreground [1][2][3][4][5]. The need for recombinant proteins with post-translational modifications has fostered the development of mammalian cell-based expression systems rather than prokaryotes and lower eukaryotes [6].…”

Section: Introductionmentioning

confidence: 99%