The response of N2 fixation and [Formula: see text] uptake to environmental conditions and nutrient enrichment experiments in three western U.S. lake systems was studied (eutrophic Clear Lake; mesotrophic Walker Lake; oligotrophic Lake Tahoe). We tested the effect of additions of bioactive trace metals molybdenum as Mo(V) and iron (Fe) as well as phosphate (P), N2 fixation, [Formula: see text], carbon (C) fixation, chlorophyll a (Chla), and bacterial cell counts under both natural conditions and in mesocosm experiments. We found distinct background N2 fixation and [Formula: see text] uptake rates: highest at Clear Lake (N2 fixation: 44.7 ± 1.8 nmol N L(-1) h(-1)), intermediate at Walker Lake (N2 fixation: 1.7 ± 1.1 nmol N L(-1) h(-1); [Formula: see text] uptake: 113 ± 37 nmol N L(-1) h(-1)), and lowest at Lake Tahoe (N2 fixation: 0.1 ± 0.07 nmol N L(-1) h(-1); [Formula: see text] uptake: 37.2 ± 10.0 nmol N L(-1) h(-1)). N2 fixation was stimulated above control values with the addition of Fe and Pin Clear Lake (up to 50 and 63%, respectively); with Mo(V), Fe, and P in Walker Lake (up to 121, 990, and 85%, respectively); and with Mo(V) and P in Lake Tahoe (up to 475 and 21%, respectively). [Formula: see text] uptake showed the highest stimulation in Lake Tahoe during September 2010, with the addition of P and Mo(V) (∼84% for both). High responses to Mo(V) additions were also observed at some sites for C fixation (Lake Tahoe: 141%), Chla (Walker Lake: 54% and Clear Lake: 102%), and bacterial cell counts (Lake Tahoe: 61%). Overall our results suggest that co-limitation of nutrients is probably a common feature in lakes, and that some trace metals may play a crucial role in limiting N2 fixation and [Formula: see text] uptake activity, though primarily in non-eutrophic lakes.