------------------- GENERAL INFORMATION ------------------- Related publication (journal paper, book, etc.): María Ortiz-Rodríguez Saurabh P. Singh, Francisco J. Cao-García, Roberto Galletto, Borja Ibarra. Regulation of Pfh1 helicase activity by nucleic acid interactions and mitochondrial SSB. Date(s) of data collection: 2021-2025 Geographic location of data collection: IMDEA Nanociencia, Madrid General description: Data reported in a scientific article -------------------------- SHARING/ACCESS INFORMATION -------------------------- Licenses/restrictions placed on the data, or limitations of reuse: N.A. Links/relationships to previous or related data sets: N.A. -------------------- DATA & FILE OVERVIEW -------------------- File list: Relationship between files: FOR ALL DATA SECTIONS: Processivity or number of nucleotides unwound is expressed in base pairs and velocities in base pairs per second. Data can be processed using Origin, Excel or related software. • 2_and_20nM  Number of raw data used for every condition. • ssDNA_2nM_Pfh1  Processivity and average translocation velocity for Pfh1 (2nM) during ssDNA translocation measured at 100, 150, 250, 375, 500, 1000 and 1500 µM ATP concentration and at 10, 15, 20 and 25 pN. • RNA-DNA_2nM_Pfh1_2nM_Rim1  Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during RNA-DNA unwinding in presence of spRim1, measured at 75 and 500 µM ATP concentration and from 5 to 11 pN. • RNA-DNA_2nM_Pfh1  Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during RNA-DNA unwinding, measured at 75 and 500 µM ATP concentration and from 5 to 11 pN. • 2nMPfh1_2nMHmtSSB  Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during DNA unwinding in presence of HmtSSB, measured at 25 and 250 µM ATP concentration and from 5 to 11 pN. • ATPgS  Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during DNA unwinding, measured at 250 µM ATPgS-50 µM ATP and concentration and 500 µM ATPgS-50 µM ATP from 5 to 11 pN. • 2nMPfh1_2nMRim1  Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during DNA unwinding in presence of spRim1, measured at 25, 50, 75, 100, 150, 250, 500 and 1000 µM ATP concentration and from 5 to 11 pN. • 20nM_Pfh1 Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (20nM) during DNA, measured at 50, 100 and 500 µM ATP concentration and from 5 to 11 pN. • 2nM_Pfh1 Number of nucleotides unwound, average unwinding velocity, rewinding velocity and their associated error for Pfh1 (2nM) during DNA unwinding, measured at 5, 25, 50, 75, 100, 150, 250, 500 and 1000 µM ATP concentration and from 5 to 11 pN. • Fits_MM  R2 obtained from the Michaelis-Menten fits for the forward and reverse velocity in absence and in presence of spRim1 and for Pfh1 translocation rate in ssDNA. Formats: .xlsx Version: Microsoft 365 -------------------------- METHODOLOGICAL INFORMATION -------------------------- Description of methods used for *collection/generation* of data: Dual-beam optical tweezers and Dual-trap optical tweezers combined with confocal microscopy Methods for *processing* the data: Matlab custom written software Software- or Instrument-specific information needed to *access/open/interpret* the data, including software and hardware version numbers: Origin and/or Excel Standards and calibration information: N.A. Environmental/experimental conditions: Helicase working buffer (described in manuscript). Data taken at room temperature Describe any quality-assurance procedures performed on the data: total number of data 2036 -------------------------- DATA-SPECIFIC INFORMATION -------------------------- Variable list, defining any abbreviations, units of measure, codes or symbols used: Processivity (nt), Av. Vel (nt/sec), Unwound bp and Error u-bp (bp), Av. Unwinding vel, Error av. Unw. Vel, Av. Rewinding vel and Error av. Rew. Vel (bp/sec)