Fatigue crack growth experiments were carried out on Ti6242 alloy with large colony size. The alloy was heat treated to provide three different lamella size; fine, coarse, and extra coarse. Tests were conducted at two temperatures, 520 °C and 595 °C, using two loading frequencies, 10 and 0.05 Hz. The latter frequency was examined with and without a 300-second hold time. All tests were performed in air environment and at a stress ratio of 0.1. This study shows that at 520 °C, the Fatigue crack growth rate (FCGR) is not significantly influenced by changes in the microstructure. For 0.05 Hz/low ΔK, however, the FCGR is higher in the fine lamellar microstructure and is accompanied by- the appearance of a plateau, which disappears in the extra large lamella microstructure. Furthermore, the addition of a 300-second hold time does not alter the crack growth rate. At 595 °C, while the general level of the FCGR is higher than that at 520 °C, the effects of loading frequency and hold time remain similar to those reported at the lower temperature. Unlike the results at 520 °C, however, the FCGR at low δK is not influenced by variations in lamellar microstructure. Under all test conditions, the fatigue process is predominantly controlled by one single mechanism associated with transcolony fracture and formation of quasi-cleavage facets. The fatigue crack growth results and the associated fracture behavior as obtained in this study are correlated to the crack-tip shear activity and transmission at the α/β interfaces. A general hypothesis accounting for the role of loading frequency, temperature, and microstructure on the observed cracking mechanisms is presented.