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Publication Date

2010-06-23

Availability

UM campus only

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PHD)

Department

Microbiology and Immunology (Medicine)

Date of Defense

2010-06-10

First Committee Member

George P. Munson

Second Committee Member

Kenneth A. Fields

Third Committee Member

Lisa R. W. Plano

Fourth Committee Member

Terace M. Fletcher

Fifth Committee Member

Timothy L. Yahr

Sixth Committee Member

Gregory Plano - Mentor

Abstract

YscD is an essential component of the plasmid pCD1-encoded type III secretion system (T3SS) of Yersinia pestis. YscD has a single transmembrane (TM) domain that connects a small N-terminal cytoplasmic region (residues 1 to 121) to a larger periplasmic region (residues 143 to 419). Deletion analyses demonstrated that both the N-terminal cytoplasmic region and the C-terminal periplasmic region are essential for YscD function. Additional studies demonstrated that a predicted cytoplasmic forkhead-associated (FHA) domain of YscD is also required for function; in contrast, a predicted periplasmic phospholipid binding (BON) domain and a putative periplasmic "ring-building" domain of YscD could be deleted with no significant effect on the T3S process. Although deletion of the putative "ring-building" domain did not disrupt T3S activity per se, the calcium-dependent regulation of the T3S apparatus was affected. The extreme C-terminal region of YscD (residues 354 to 419) was essential for secretion activity and had a strong dominant negative effect on the T3S process when exported to the periplasm of the wild type parent strain. Finally, replacement of the YscD TM domain with a TM domain of dissimilar sequence had no effect on the T3S process, indicating that the TM domain has no sequence-specific function in the assembly or function of the T3SS.

Keywords

Bacteria; Microbiology

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