The influence of surface heat treatment using laser radiation on the fatigue strength and corresponding microstructural evolution of AISI 4140 alloy steel was investigated in this research. The AISI 4140 alloy steel was radiated by a diode laser to give surface temperatures in the range between 600 and 800°C, and subsequently underwent vibration peening. The fatigue behavior of surface-treated specimens was examined using a giga-cycle ultrasonic fatigue test, and it was compared with that of non-treated and only-peened specimens. Fatigue fractured surfaces and microstructural evolution with respect to the laser treatment temperatures were investigated using an optical microscope. Hardness distribution was measured using Vickers micro-hardness. Higher laser temperature resulted in higher fatigue strength, attributed to the phase transformation.