The Aspergillus nidulans hapC gene was expressed as a fusion protein with MalE or glutathione-S-transferase (GST) in Escherichia coli, and used for the purification of HapC and the preparation of anti-HapC antiserum. The CCAAT-binding factor AnCP/AnCF contains a component with an approximate molecular mass of 32 kDa that cross-reacts with the antibody. The MalE-HapC fusion protein was able to replace authentic HapC in AnCP when incubated under appropriate conditions. Furthermore, reconstitution experiments with recombinant HapC, yHAP2 and yHAP5 polypeptides showed that all three polypeptides were required for the assembly of a complex capable of binding to CCAAT-containing taaG2 promoter DNA. The relationship between AnCP/AnCF and the Saccharomyces cerevisiae HAP complex is discussed.

• PMID: 9529521
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Reference Type

Comparative Study | Journal Article

Authors

Kato M, Aoyama A, Naruse F, Tateyama Y, Hayashi K, Miyazaki M, Papagiannopoulos P, Davis MA, Hynes MJ, Kobayashi T, ... Show all

Primary Lit For

CCAAT-binding factor complex

Additional Lit For

HAP2 | HAP4 | HAP3 | HAP5

Review For