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CotC-CotU Heterodimerization during Assembly of the Bacillus subtilis Spore Coat

Rachele Isticato,¹ Assunta Pelosi,¹ Rita Zilhão,^2,3, Loredana Baccigalupi,¹ Adriano O. Henriques,³ Maurilio De Felice,¹ and Ezio Ricca^1*

Dipartimento di Biologia Strutturale e Funzionale, Università Federico II, Naples, Italy,¹ Departmento de Biologia Vegetal, Universidade de Lisboa, Lisbon, Portugal,² Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras, Portugal³

Received 3 September 2007/ Accepted 25 November 2007

We report evidence that CotC and CotU, two previously identified components of the Bacillus subtilis spore coat, are produced concurrently in the mother cell chamber of the sporulating cell under the control of ^K and GerE and immediately assembled around the forming spore. In the coat, the two proteins interact to form a coat component of 23 kDa. The CotU-CotC interaction was not detected in two heterologous hosts, suggesting that it occurs only in B. subtilis. Monomeric forms of both CotU and CotC failed to be assembled at the surface of the developing spore and accumulated in the mother cell compartment of cells mutant for cotE. In contrast, neither CotU nor CotC accumulated in the mother cell compartment of cells mutant for cotH. These results suggest that CotH is required to protect both CotU and CotC in the mother cell compartment of the sporangium and that CotE is needed to allow their assembly and subsequent interaction at the spore surface.

Published ahead of print on 7 December 2007.

Present address: Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisbon, Portugal.