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Series GSE102981 Query DataSets for GSE102981
Status Public on Aug 23, 2018
Title Sequence context controls the DNA binding and processivity of the T7 DNA primase
Platform organism synthetic construct
Sample organism Escherichia phage T7
Experiment type Other
Summary Primases are key enzymes involved in DNA replication. They act on single-stranded DNA, and catalyze the synthesis of short RNA primers used by DNA polymerases. Here, we investigate the DNA-binding and functional activity of the bacteriophage T7 primase using a new workflow called High-Throughput Primase Profiling (HTPP). Using a unique combination of high-throughput binding assays and biochemical analyses, HTPP reveals a complex landscape of binding specificity and functional activity for the T7 primase, determined by sequences flanking the primase recognition site. We identified specific features, such at G/T-rich flanks, which in-crease primase-DNA binding up to 10-fold and, surprisingly, also increase the length of newly formed RNA (2-3 fold). To our knowledge, variability in primer length has not been reported for this primase. We expect that applying HTPP to additional enzymes will reveal new insights into the effects of DNA sequence composition on the DNA recognition and functional activity of primases.
 
Overall design HTPP (High-Throughput Primase Profiling) experiments were performed for recombinant T7 primase residue: 1-271. Briefly, the HTPP involved binding of his-tagged primase to single-stranded 180K Agilent microarrays in order to determine their binding specificity in different artificial context. Each sequence represented on the array is present in 6 replicate spots. We report the signal intensity for each spot.
 
Contributor(s) Gordan R, Afek A
Citation(s) 30428370
Submission date Aug 23, 2017
Last update date Jul 25, 2021
Contact name Raluca Gordan
E-mail(s) raluca.gordan@duke.edu
Organization name Duke University
Department Center for Genomic and Computational Biology
Street address 101 Science Dr, CIEMAS 2179
City Durham
State/province NC
ZIP/Postal code 27708
Country USA
 
Platforms (1)
GPL23947 Duke/RG_AA_T7Primase_v1
Samples (1)
GSM2752104 T7 primase (residue: 1-271) at 5 uM concentration
Relations
BioProject PRJNA399696

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE102981_RAW.tar 11.2 Mb (http)(custom) TAR (of TXT)
GSE102981_T7primase_5uM_Mg_6replicates_and_median.xlsx 2.3 Mb (ftp)(http) XLSX
Processed data included within Sample table
Processed data are available on Series record

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