Knockout of the Ribonuclease Inhibitor Gene Leaves Human Cells Vulnerable to Secretory Ribonucleases.
Ribonuclease inhibitor (RNH1) is a cytosolic protein that binds with femtomolar affinity to human ribonuclease 1 (RNase 1) and homologous secretory ribonucleases. RNH1 contains 32 cysteine residues and has been implicated as an antioxidant. Here, we use CRISPR-Cas9 to knock out RNH1 in HeLa cells. We find that cellular RNH1 affords marked protection from the lethal ribonucleolytic activity of RNase 1 but not from oxidants. We conclude that RNH1 protects cytosolic RNA from invading ribonucleases.
Graduate Program in Cellular & Molecular Biology, University of Wisconsin-Madison , 1525 Linden Drive, Madison, Wisconsin 53706, United States.
Sydney P
SP
Thomas
Center for Genome Engineering, Institute for Basic Science , Seoul 08826, Republic of Korea.
Eunji
E
Kim
Center for Genome Engineering, Institute for Basic Science , Seoul 08826, Republic of Korea.
Jin-Soo
JS
Kim
Department of Biochemistry, University of Wisconsin-Madison , 433 Babcock Drive, Madison, Wisconsin 53706, United States.|Department of Chemistry, University of Wisconsin-Madison , 1101 University Avenue, Madison, Wisconsin 53706, United States.
Ronald T
RT
Raines
D064113:CRISPR-Cas Systems; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*
D002352:Carrier Proteins; D016877:Oxidants; D012334:RNA, Neoplasm; C501467:RNH1 protein, human; D006861:Hydrogen Peroxide; D012259:Ribonuclease, Pancreatic