Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila

A large number of proteins transferred by the Legionella pneumophila Dot/Icm system have been identified by various strategies. With no exceptions, these strategies are based on one or more characteristics associated with the tested proteins. Given the high level of diversity exhibited by the identi...

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Main Authors:	Zhu, Wenhan, Banga, Simran, Tan, Yunhao, Zheng, Cheng, Stephenson, Robert, Gately, Jonathan, Luo, Zhao-Qing
Format:	Online
Language:	English
Published:	Public Library of Science 2011
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3052360/

id	pubmed-3052360
recordtype	oai_dc
spelling	pubmed-30523602011-03-15 Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila Zhu, Wenhan Banga, Simran Tan, Yunhao Zheng, Cheng Stephenson, Robert Gately, Jonathan Luo, Zhao-Qing Research Article A large number of proteins transferred by the Legionella pneumophila Dot/Icm system have been identified by various strategies. With no exceptions, these strategies are based on one or more characteristics associated with the tested proteins. Given the high level of diversity exhibited by the identified proteins, it is possible that some substrates have been missed in these screenings. In this study, we took a systematic method to survey the L. pneumophila genome by testing hypothetical orfs larger than 300 base pairs for Dot/Icm-dependent translocation. 798 of the 832 analyzed orfs were successfully fused to the carboxyl end of β-lactamase. The transfer of the fusions into mammalian cells was determined using the β-lactamase reporter substrate CCF4-AM. These efforts led to the identification of 164 proteins positive in translocation. Among these, 70 proteins are novel substrates of the Dot/Icm system. These results brought the total number of experimentally confirmed Dot/Icm substrates to 275. Sequence analysis of the C-termini of these identified proteins revealed that Lpg2844, which contains few features known to be important for Dot/Icm-dependent protein transfer can be translocated at a high efficiency. Thus, our efforts have identified a large number of novel substrates of the Dot/Icm system and have revealed the diverse features recognizable by this protein transporter. Public Library of Science 2011-03-09 /pmc/articles/PMC3052360/ /pubmed/21408005 http://dx.doi.org/10.1371/journal.pone.0017638 Text en Luo et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Zhu, Wenhan Banga, Simran Tan, Yunhao Zheng, Cheng Stephenson, Robert Gately, Jonathan Luo, Zhao-Qing
spellingShingle	Zhu, Wenhan Banga, Simran Tan, Yunhao Zheng, Cheng Stephenson, Robert Gately, Jonathan Luo, Zhao-Qing Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
author_facet	Zhu, Wenhan Banga, Simran Tan, Yunhao Zheng, Cheng Stephenson, Robert Gately, Jonathan Luo, Zhao-Qing
author_sort	Zhu, Wenhan
title	Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
title_short	Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
title_full	Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
title_fullStr	Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
title_full_unstemmed	Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila
title_sort	comprehensive identification of protein substrates of the dot/icm type iv transporter of legionella pneumophila
description	A large number of proteins transferred by the Legionella pneumophila Dot/Icm system have been identified by various strategies. With no exceptions, these strategies are based on one or more characteristics associated with the tested proteins. Given the high level of diversity exhibited by the identified proteins, it is possible that some substrates have been missed in these screenings. In this study, we took a systematic method to survey the L. pneumophila genome by testing hypothetical orfs larger than 300 base pairs for Dot/Icm-dependent translocation. 798 of the 832 analyzed orfs were successfully fused to the carboxyl end of β-lactamase. The transfer of the fusions into mammalian cells was determined using the β-lactamase reporter substrate CCF4-AM. These efforts led to the identification of 164 proteins positive in translocation. Among these, 70 proteins are novel substrates of the Dot/Icm system. These results brought the total number of experimentally confirmed Dot/Icm substrates to 275. Sequence analysis of the C-termini of these identified proteins revealed that Lpg2844, which contains few features known to be important for Dot/Icm-dependent protein transfer can be translocated at a high efficiency. Thus, our efforts have identified a large number of novel substrates of the Dot/Icm system and have revealed the diverse features recognizable by this protein transporter.
publisher	Public Library of Science
publishDate	2011
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3052360/
_version_	1611444026498613248

Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila

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