Eukaryotic cells can sense glucose and evoke signaling pathways to regulate growth and development. An immediate response to glucose is the expression of a set of genes mediated by cis carbohydrate response elements (ChoRE) and their associated transcription factors MondoA and Max-like protein X (MLX). Thioredoxin interacting protein (TXNIP), the product of an immediate glucose response gene TXNIP, functions as a negative regulator for glucose uptake, and its expression is dysregulated in diabetes and cancer. We have observed that the ChoRE cis regulatory element is duplicated during vertebrate evolution, with one ChoRE in fish and two in mammals. In mammalian cells, both ChoREs are required for an optimal glucose response. With assistance by nuclear factor Y (NF-Y), MondoA/MLX complex is recruited to TXNIP promoter upon glucose stimulation, which in turn recruits general transcription factors and RNA polymerases to initiate gene transcription. In addition to glucose or its derived metabolites, MondoA/MLX activity and TXNIP expression is tightly correlated with status of mitochondrial oxidative phosphorylation (OXPHOS), and inhibition of OXPHOS by drugs such as metformin can dramatically repress TXNIP transcription by inducing glycolytic flux. Moreover, we have discovered that the expression of TXNIP is induced by an array of adenosine-containing molecules, and these molecules function as amplifiers of glucose signaling. Thus, MondoA/MLX complex serves as a hub integrating diverse upstream signals and a master regulator of glucose homeostasis.