Vam6, Gtr1/2, and Ego1/3 are required for leucine-dependent TorC1 kinase activation which is central to nitrogen-responsive regulation. However, Gln3, a nitrogen-responsive transcription activator, does not respond to leucine-dependent TorC1 activation. In nitrogen excess, Gln3 is cytoplasmic and Gln3-mediated transcription minimal, whereas in nitrogen limitation, starvation, or following rapamycin treatment, Gln3 is nuclear and transcription greatly increased. Increasing evidence demonstrates nitrogen-responsive intracellular Gln3 localization is subject to multiple modes of regulation. To ascertain whether the Vam6, Gtr-Ego complexes participate in the regulation of Gln3, we determined the requirements of the above proteins for nuclear localization and cytoplasmic sequestration of Gln3 in response to nitrogen excess, starvation or limitation. We show that Gln3 is sequestered in the cytoplasm of vam6Δ, gtr1Δ, gtr2Δ, ego1Δ and ego3Δ either long-term in logarithmically glutamine-grown cells or short-term a...