TY - JOUR T1 - Genome-wide analysis of in vivo CcpA binding with and without its key co-factor HPr in the major human pathogen group A <em>Streptococcus</em> JF - bioRxiv DO - 10.1101/2020.08.28.272682 SP - 2020.08.28.272682 AU - Sruti DebRoy AU - Victor Aliaga Tobar AU - Gabriel Galvez AU - Srishtee Arora AU - Xiaowen Liang AU - Nicola Horstmann AU - Vinicius Maracaja-Coutinho AU - Mauricio Latorre AU - Magnus Hook AU - Anthony R. Flores AU - Samuel A. Shelburne Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/08/28/2020.08.28.272682.abstract N2 - Catabolite control protein A (CcpA) is a master regulator of carbon source utilization and contributes to the virulence of numerous medically important Gram-positive bacteria. Most functional assessments of CcpA, including interaction with its key co-factor HPr, have been performed in non-pathogenic bacteria. In this study we aimed to identify the in vivo DNA binding profile of CcpA and assess the extent to which HPr is required for CcpA-mediated regulation and DNA binding in the major human pathogen group A Streptococcus (GAS). Using a combination RNAseq/ChIPseq approach, we found that CcpA affects transcript levels of 514 of 1667 GAS genes (31%) whereas direct DNA binding was identified for 105 GAS genes. Three of the directly regulated genes encode the key GAS virulence factors Streptolysin S, PrtS (IL-8 degrading proteinase), and SpeB (cysteine protease). Mutating CcpA Val301 to Ala (strain 2221-CcpA-V301A) abolished interaction between CcpA and HPr and impacted the transcript levels of 205 genes (40%) in the total CcpA regulon. By ChIPseq analysis, CcpAV301A bound to DNA from 74% of genes bound by wild-type CcpA, but generally with lower affinity. These data delineate the direct CcpA regulon and clarify the HPr-dependent and independent activities of CcpA in a key pathogenic bacterium.Data sharing and data availability The data that support the findings of this study are available from the corresponding author upon reasonable request.Competing Interest StatementThe authors have declared no competing interest. ER -