Source Data for "Defining the ATPome Reveals Cross-Optimization of Metabolic Pathways"

Disrupted energy metabolism drives cell dysfunction and disease,
but approaches to increase or preserve ATP are lacking. To generate a
comprehensive metabolic map of genes and pathways that regulate cellular ATP — the
ATPome — we conducted a genome-wide CRISPR interference/activation screen
integrated with an ATP-biosensor. We show
that ATP level is modulated by distinct mechanisms that promote energy
production or inhibit consumption. Surprisingly, in our system HK2 is the greatest
ATP consumer , indicating energy failure may not be a general deficiency in
producing ATP, but rather failure to recoup the ATP cost of glycolysis and diversion
of glucose metabolites to the pentose phosphate pathway. We identify systems-level
reciprocal inhibition between the HIF1 pathway and mitochondria;
glycolysis-promoting enzymes inhibit respiration even when there is no
glycolytic ATP production, and vice versa. Consequently, suppressing
alternative metabolism modes paradoxically increases energy levels under
substrate restriction. This work reveals mechanisms of metabolic control, and
identifies therapeutic targets to correct energy failure. This Source Data file contains source data and statistics for the figures of this manuscript.