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Signals that facilitate and coordinate the cross talk as the bronchial forming canalicular stage transitions to construction of air-exchanging capillary-alveoli niche in the alveolar stage are poorly understood. Expressed within this decisive region, levels of aminoacyl-tRNA synthetase complex-interacting multifunctional protein 1 (AIMP1) inversely correlate with the maturation of the lung. The present study addresses the role of AIMP1 in lung development through the generation and characterization of Aimp1<jats:sup>\u2212\/\u2212<\/jats:sup>mutant mice. Mating of Aimp1<jats:sup>+\/\u2212<\/jats:sup>produced offspring in expected Mendelian ratios throughout embryonic development. However, newborn Aimp1<jats:sup>\u2212\/\u2212<\/jats:sup>pups exhibited neonatal lethality with mild cyanosis. Imaging both structure and ultrastructure of Aimp1<jats:sup>\u2212\/\u2212<\/jats:sup>lungs showed disorganized bronchial epithelium, decreased type I but not type II cell differentiation, increased distal vessels, and disruption of E-cadherin deposition in cell-cell junctions. Supporting the in vivo findings of disrupted epithelial cell-cell junctions, in vitro biochemical experiments show that a portion of AIMP1 binds to phosphoinositides, the lipid anchor of proteins that have a fundamental role in both cellular membrane and actin cytoskeleton organization; a dramatic disruption in F-actin cytoskeleton was observed in Aimp1<jats:sup>\u2212\/\u2212<\/jats:sup>mouse embryonic fibroblasts. Such observed structural defects may lead to disrupted cell-cell boundaries. 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