The order must be K2→K1, since the permanently active K1 allele (K1a) is able to propagate the signal onward even when its upstream activator K2 is inactive (K2i). The reverse order would have resulted in a failure to signal (K1a→K2i), since the permanently active K1a kinase would be attempting to activate a dead K2i kinase.
Explanation:
You characterize a double mutant cell that contains K2 with type I mutation and K1 with type II mutation. You observe that the response is seen even when no extracellular signal is provided. In the normal pathway, i f K1 activat es K2, we expect t his combinat ion of two m utants to show no response with or without ext racell ular signal. This is because no matt er how active K1 i s, it would be unable to act ivate a mutant K2 that i s an activit y defi cient. If we reverse the order, K2 activating K1, the above observati on is valid. Therefore, in the normal signaling pathway, K2 activates K1.A: land and D: fresh water