The centromere is the chromosomal site of kinetochore assembly, defined by the histone H3 variant CENP-A. In each cell cycle, the assembly and maintenance of CENP-A is functionally critical for chromosome segregation. In Drosophila male meiosis, CID (fly CENP-A) is assembled in two phases: prophase of meiosis I and postmeiosis II. Here, we investigate the dynamics of the assembly components CAL1 and CENP-C in prophase I and determine the requirements for the second assembly phase. In early prophase I, CENP-C functions with CAL1 to maintain the centromere. In late prophase I, CAL1 is undetectable at centromeres and CENP-C is not critical for centromere maintenance. Instead, CENP-C is crucial for meiotic kinetochore recruitment and function. This CENP-C pool also functions in CID assembly postmeiosis II, which is independent of CAL1. In addition to different functional pools of CENP-C, distinct pools of the CID protein persist in the male germline, and the synthesis of each pool is uncoupled from its cell cycle deposition timing.
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