Structural remodeling of Coxiella burnetii during its biphasic developmental cycle revealed by cryo-electron tomography

Abstract

Coxiella burnetii is an obligate zoonotic bacterium that targets macrophages to cause a disease known as Q fever. It has a biphasic developmental lifecycle where the extracellular and metabolically inactive small cell variant (SCV) transforms, under host acidic environment, into the vegetative large cell variant (LCV). However, the details about the morphological and structural changes that accompany this biphasic cycle are still lacking. Here, we used cryo-electron tomography to image the different cell variants of C. burnetii grown either under axenic conditions in different pH or purified directly from host cells revealing the major developmental, morphological and structural transitions. We show that SCVs are characterized by equidistant stacks of inner membrane that presumably allow a smooth transition to LCV, a transition coupled with the expression of the Dot/Icm type IVB secretion system (T4BSS). A class of T4BSS particles were associated with extracellular densities including a tubular structure possibly involved in host interaction or effector delivery. Also, SCVs and cells in the transition state contained spherical multilayered membrane structures of different sizes and locations suggesting that they are not related to a sporulation process as once assumed.

Structure and Function of the Dot/Icm T4SS

“…In situ (8)(9)(10)(11)(12)(13) and in vitro (14)(15)(16)(17) structural studies of T4SSs have revealed their complexity and elegance. Our early cryo-electron tomography (cryo-ET) studies (8,10), and those of others (12,13), revealed the overall architecture of the intact Dot/Icm T4SS in situ and began to dissect the molecular organization of this complex (10,12).…”