BipA is a conserved ribosome-associated GTPase that supports bacterial stress adaptation. It participates in 50S ribosome assembly and envelope stress responses; however, it is not known whether both functions depend on its GTPase activity. In this study, we investigated Escherichia coli strains carrying chromosomal D74A or N128D substitutions in the conserved GTPase motifs of BipA. Both mutants displayed severe cold-sensitive growth defects that were similar to those of the ΔbipA strain. At low temperatures, 50S ribosome assembly was defective and immature ribosomal particles accumulated. Ribosome-binding analysis revealed that the mutant BipA had a lower association with ribosomal fractions. The mutants exhibited altered lipopolysaccharide profiles and envelope phenotypes, including increased capsule production, reduced biofilm formation and motility, and increased cell width and length. These phenotypes were most notable at low temperatures, indicating activation of the Rcs (Regulator of Capsule Synthesis) phosphorelay. We next examined the effects of BipA abundance on cellular physiology. BipA expression was influenced by both start-codon identity and the downstream coding context. BipA overexpression impaired growth and induced pronounced cell elongation at both optimal and low temperatures, indicating that proper control of BipA abundance is important for cellular homeostasis. Furthermore, the increased susceptibility of the ΔbipA strain to ampicillin at low temperatures suggested an association between BipA and peptidoglycan homeostasis. Together, these findings suggest that BipA GTPase function is required to maintain ribosome biogenesis and envelope homeostasis during cold adaptation, and that proper control of BipA abundance is also important for maintaining cellular physiology.