國立虎尾科技大學 |

Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana.

紀錄類型:	書目-語言資料,手稿 : Monograph/item
正題名/作者:	Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana./
作者:	Groves, Norman R.
面頁冊數:	1 online resource (230 pages)
附註:	Source: Dissertations Abstracts International, Volume: 81-08, Section: B.
Contained By:	Dissertations Abstracts International81-08B.
標題:	Botany. -
電子資源:	click for full text (PQDT)
ISBN:	9781392345337

Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana.
Groves, Norman R.

Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana. - 1 online resource (230 pages)

Source: Dissertations Abstracts International, Volume: 81-08, Section: B.

Thesis (Ph.D.)--The Ohio State University, 2019.

Includes bibliographical references

Proteins residing at the inner nuclear membrane (INM) in animals and yeast play a variety of roles within the cell, including nuclear organization, nuclear movement and chromatin organization. INM proteins in animals and yeast have been shown to have a variety of topologies and structures. Over 100 nuclear envelope transmembrane proteins (NETs) have been shown to be enriched at nuclear membranes, but have yet to be tested for INM localization. Amongst known INM proteins in animals and yeast, only SUN proteins are encoded for in plant genomes. SUN proteins form the INM component of linker of nucleoskeleton and cytoskeleton (LINC) complexes and are conserved from animals and yeast to plants. Besides SUN proteins, cation channels have been identified that localize to the INM, in the form of Medicago truncatula DMI1 and CNGC15. Given the lack of homologs of mammalian INM proteins in plants, proteins likely exist that are structurally divergent from animal and yeast INM proteins, but functionally serve similar roles within the cell.In both plants and animals, protein trafficking to the INM is poorly understood relative to other protein targeting pathways. One model for protein trafficking to the INM is transport factor-mediated trafficking, in which nuclear localization signals (NLSs) are necessary for localization at the INM. To determine if such a pathway to the INM exists in plants, I fused an NLS to a tail-anchored ER protein, PICL, and determined that a GFP-tagged, NLS-fused PICL is enriched at the NE in Nicotiana benthamiana leaf epidermal cells. Localization at the INM compared to outer nuclear membrane (ONM) was resolved using Airyscan sub-diffraction limited confocal microscopy, and NLS-fused PICL was shown to access the INM. Nine monopartite and bipartite NLSs were tested for their sufficiency to enrich a chimeric membrane protein, Heh2linker-TMD, at the NE. All nine NLSs were sufficient for enrichment of the chimeric membrane at the NE, and a monopartite and bipartite NLS were each shown to be sufficient for localization to the INM. This data is the first evidence that an NLS-mediated pathway to the INM exists in plants. Presence of an NLS and transmembrane domain (TMD) was used as criterium to identify novel INM proteins in plants. The Arabidopsis thaliana proteome was queried for presence of both domains, resulting in a shortlist of 32 putative inner nuclear envelope proteins (PINEs). A secondary search for Arabidopsis homologs of mammalian nuclear envelope transmembrane proteins (NETs) yielded 16 proteins, Arabidopsis SUN1 among them. The results of these queries are presented here.While the INM component of LINC complexes, SUN proteins, are conserved from animals and yeast to plants, the ONM components of plant LINC complexes has diverged in plants. Those proteins, KASH proteins, are defined by a conserved C-terminus consisting of a TMD and NE luminal tail. KASH proteins vary in their cytoplasmic domains, which interact directly or indirectly with the cytoskeleton. While no sequence homologs of animal KASH proteins exist, functional homologs have been identified in Arabidopsis. The plant KASH proteins WIPs, SINE1 and SINE2 have been shown to interact with plant SUN proteins, confirming the presence of plant LINC complexes. WIPs and SINEs have been shown to be required for nuclear shape determination, nuclear movement and nuclear positioning. The proteins that interact with plant KASH proteins largely remain unidentified. Here, I conducted a split ubiquitin yeast 2-hybrid screen of the Arabidopsis genome to identify the cytoplasmic components of plant LINC complexes. The work presented here expands our knowledge or how protein trafficking to the INM occurs in plants, and presents candidates for NE-associated proteins at both the INM and ONM. This work will serve as a platform to increase our understanding of the plant nuclear envelope, and further understand the differences between the plant and animal nuclear envelope.

Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2024

Mode of access: World Wide Web

ISBN: 9781392345337Subjects--Topical Terms:

599558
Botany.
Subjects--Index Terms:

Protein traffickingIndex Terms--Genre/Form:

554714
Electronic books.

Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana.
LDR:05632ntm a22004457 4500 001 1148109
005 20240916070001.5
006 m o d
007 cr bn ---uuuuu
008 250605s2019 xx obm 000 0 eng d
020 $a 9781392345337
035 $a (MiAaPQ)AAI27712047
035 $a (MiAaPQ)OhioLINKosu1557145385558223
035 $a AAI27712047
040 $a MiAaPQ $b eng $c MiAaPQ $d NTU
100 1 $a Groves, Norman R. $3 1474004
245 1 0 $a Determining Features Sufficient for Protein Trafficking to the Plant Inner Nuclear Membrane and Identification of Putative Nuclear Envelope-associated Proteins in Arabidopsis thaliana.
264 0 $c 2019
300 $a 1 online resource (230 pages)
336 $a text $b txt $2 rdacontent
337 $a computer $b c $2 rdamedia
338 $a online resource $b cr $2 rdacarrier
500 $a Source: Dissertations Abstracts International, Volume: 81-08, Section: B.
500 $a Advisor: Meier , Iris.
502 $a Thesis (Ph.D.)--The Ohio State University, 2019.
504 $a Includes bibliographical references
520 $a Proteins residing at the inner nuclear membrane (INM) in animals and yeast play a variety of roles within the cell, including nuclear organization, nuclear movement and chromatin organization. INM proteins in animals and yeast have been shown to have a variety of topologies and structures. Over 100 nuclear envelope transmembrane proteins (NETs) have been shown to be enriched at nuclear membranes, but have yet to be tested for INM localization. Amongst known INM proteins in animals and yeast, only SUN proteins are encoded for in plant genomes. SUN proteins form the INM component of linker of nucleoskeleton and cytoskeleton (LINC) complexes and are conserved from animals and yeast to plants. Besides SUN proteins, cation channels have been identified that localize to the INM, in the form of Medicago truncatula DMI1 and CNGC15. Given the lack of homologs of mammalian INM proteins in plants, proteins likely exist that are structurally divergent from animal and yeast INM proteins, but functionally serve similar roles within the cell.In both plants and animals, protein trafficking to the INM is poorly understood relative to other protein targeting pathways. One model for protein trafficking to the INM is transport factor-mediated trafficking, in which nuclear localization signals (NLSs) are necessary for localization at the INM. To determine if such a pathway to the INM exists in plants, I fused an NLS to a tail-anchored ER protein, PICL, and determined that a GFP-tagged, NLS-fused PICL is enriched at the NE in Nicotiana benthamiana leaf epidermal cells. Localization at the INM compared to outer nuclear membrane (ONM) was resolved using Airyscan sub-diffraction limited confocal microscopy, and NLS-fused PICL was shown to access the INM. Nine monopartite and bipartite NLSs were tested for their sufficiency to enrich a chimeric membrane protein, Heh2linker-TMD, at the NE. All nine NLSs were sufficient for enrichment of the chimeric membrane at the NE, and a monopartite and bipartite NLS were each shown to be sufficient for localization to the INM. This data is the first evidence that an NLS-mediated pathway to the INM exists in plants. Presence of an NLS and transmembrane domain (TMD) was used as criterium to identify novel INM proteins in plants. The Arabidopsis thaliana proteome was queried for presence of both domains, resulting in a shortlist of 32 putative inner nuclear envelope proteins (PINEs). A secondary search for Arabidopsis homologs of mammalian nuclear envelope transmembrane proteins (NETs) yielded 16 proteins, Arabidopsis SUN1 among them. The results of these queries are presented here.While the INM component of LINC complexes, SUN proteins, are conserved from animals and yeast to plants, the ONM components of plant LINC complexes has diverged in plants. Those proteins, KASH proteins, are defined by a conserved C-terminus consisting of a TMD and NE luminal tail. KASH proteins vary in their cytoplasmic domains, which interact directly or indirectly with the cytoskeleton. While no sequence homologs of animal KASH proteins exist, functional homologs have been identified in Arabidopsis. The plant KASH proteins WIPs, SINE1 and SINE2 have been shown to interact with plant SUN proteins, confirming the presence of plant LINC complexes. WIPs and SINEs have been shown to be required for nuclear shape determination, nuclear movement and nuclear positioning. The proteins that interact with plant KASH proteins largely remain unidentified. Here, I conducted a split ubiquitin yeast 2-hybrid screen of the Arabidopsis genome to identify the cytoplasmic components of plant LINC complexes. The work presented here expands our knowledge or how protein trafficking to the INM occurs in plants, and presents candidates for NE-associated proteins at both the INM and ONM. This work will serve as a platform to increase our understanding of the plant nuclear envelope, and further understand the differences between the plant and animal nuclear envelope.
533 $a Electronic reproduction. $b Ann Arbor, Mich. : $c ProQuest, $d 2024
538 $a Mode of access: World Wide Web
650 4 $a Botany. $3 599558
650 4 $a Molecular biology. $3 583443
650 4 $a Plant biology. $3 1190898
650 4 $a Plant sciences. $3 1179743
650 4 $a Cellular biology. $3 1148666
653 $a Protein trafficking
653 $a Tail-anchored protein
653 $a Nuclear Localization signal
653 $a Plant
653 $a Inner Nuclear Membrane
653 $a Outer Nuclear Membrane
653 $a KASH
655 7 $a Electronic books. $2 local $3 554714
690 $a 0479
690 $a 0309
690 $a 0307
690 $a 0379
710 2 $a The Ohio State University. $b Molecular Genetics. $3 1474005
710 2 $a ProQuest Information and Learning Co. $3 1178819
773 0 $t Dissertations Abstracts International $g 81-08B.
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27712047 $z click for full text (PQDT)