02bf02a07651fb198f4ee224d0a35432 26TV17_Viruses_Martin-Hernandez_pub.pdf 7ebbe3a5529a36f12a971314cd6b94061786e09b 26TV17_Viruses_Martin-Hernandez_pub.pdf 0f04e36087cf910cd5462408225c8d7fa9b429851eba7eead7cc25094431569f 26TV17_Viruses_Martin-Hernandez_pub.pdf Title: The Melon Sterol Transporter Niemann-Pick C1 Protein Is a New Interactor of Cucumber mosaic virus Movement Protein Subject: Plant viruses need to use many host factors to establish infection. During the viral cycle, intracellular transport is fundamental to reach the plasmodesmata to enable cell-to-cell transport. Cucumovirus CMV (cucumber mosaic virus, CMV) can infect plants from most economically important crops. To identify additional host proteins involved in CMV movement in melon, we used the MP as a bait to screen a Yeast two-hybrid cDNA library from CMV-infected plants and identified a Niemann-Pick C1 (NPC1) protein as a novel MP interactor. NPC1 is a transmembrane protein involved in cholesterol transport in animal cells, but also in the infection by several viruses of different families. The identified clone from the melon NPC1 gene spans from exons 25 to 28 and includes two introns. Notably, deletion of the two introns and exon 28 does not impair the interaction capacity of the remaining peptide. The identified CmNPC1 gene maps to chromosome 11. In addition, the melon genome encodes a second copy of NPC1 in chromosome 7 (CmNPC1-C7), highly similar. Functional assays revealed that the interaction domain of CmNPC1-C7 also interacts with CMV MP, suggesting that both genes could have a role in CMV infection. This study represents the first report linking NPC1 to the infection process of a plant virus, expanding our understanding of plant–virus interactions. Keywords: Niemann-Pick C1; Cucumber mosaic virus; movement protein; interactor domain; virus infection; melon Author: Núria Real, Irene Villar, Bin Liu, Manale Gajjout, Weina Hou and Ana Montserrat Martín-Hernández Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Wed May 20 11:02:04 2026 CEST ModDate: Wed May 20 12:41:08 2026 CEST Custom Metadata: yes Metadata Stream: yes Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4318318 bytes Optimized: yes PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZDVYOU+VnURWPalladioL Type 1 Custom yes yes yes 521 0 LJPQWA+URWPalladioL-Roma Type 1 Custom yes yes yes 525 0 FAQQYU+URWPalladioL-Bold Type 1 Custom yes yes yes 528 0 CRZHIK+URWPalladioL-Ital Type 1 Custom yes yes yes 531 0 QRZDHX+URWPalladioL-BoldItal Type 1 Custom yes yes yes 534 0 VIWIOF+CMSY10 Type 1 Builtin yes yes yes 267 0 ZXDKIP+EURM10 Type 1 Builtin yes yes yes 270 0 HMFDDK+PalatinoLinotype TrueType WinAnsi yes yes no 176 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-10-16 03:56:06 CEST RepresentationInformation: 26TV17_Viruses_Martin-Hernandez_pub.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-10-15 15:49:08 CEST Size: 4318318 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Linearized PDF PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Plant Material, Viral Strains and Yeast Strains Item: Title: Plasmid Construction Item: Title: RNA Isolation Item: Title: Library Construction Item: Title: Bait Vector pGBKT7 Testing and Protein Expression Item: Title: Y2H Screening of Proteins Interacting with CMV-FNY MP Item: Title: One-by-One Y2H Item: Title: Transient Expression in Nicotiana benthamiana Item: Title: Confocal Laser Scanning Microscopy Item: Title: Co-Immunoprecipitation (Co-IP) Item: Title: Bioinformatic Analysis of CmNPC1 Item: Title: Structural modeling and interfacial analysis Item: Title: Results Children: Item: Title: Screening of a Melon cDNA Library Against CMV Movement Protein Item: Title: A Niemman’s Pick C1 Protein Interacts with CMV-MP Item: Title: Structure of the CmNPC1 ID-C11 Item: Title: Introns and Exon 28 Are Dispensable for the Interaction with CMV-MPs Item: Title: Structure of the Melon NPC1 Gene Item: Title: CmNPC1-C7 Interacts with CMV MP Item: Title: Discussion Item: Title: References Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: FAQQYU+URWPalladioL-Bold FontSubset: true FirstChar: 2 LastChar: 250 FontDescriptor: FontName: FAQQYU+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: CRZHIK+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 150 FontDescriptor: FontName: CRZHIK+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: QRZDHX+URWPalladioL-BoldItal FontSubset: true FirstChar: 67 LastChar: 118 FontDescriptor: FontName: QRZDHX+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: ZDVYOU+VnURWPalladioL FontSubset: true FirstChar: 160 LastChar: 230 FontDescriptor: FontName: ZDVYOU+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: VIWIOF+CMSY10 FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: VIWIOF+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true ToUnicode: true Font: BaseFont: LJPQWA+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: LJPQWA+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: ZXDKIP+EURM10 FontSubset: true FirstChar: 11 LastChar: 22 FontDescriptor: FontName: ZXDKIP+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true ToUnicode: true XMP: application/pdf Núria Real, Irene Villar, Bin Liu, Manale Gajjout, Weina Hou and Ana Montserrat Martín-Hernández Plant viruses need to use many host factors to establish infection. During the viral cycle, intracellular transport is fundamental to reach the plasmodesmata to enable cell-to-cell transport. Cucumovirus CMV (cucumber mosaic virus, CMV) can infect plants from most economically important crops. To identify additional host proteins involved in CMV movement in melon, we used the MP as a bait to screen a Yeast two-hybrid cDNA library from CMV-infected plants and identified a Niemann-Pick C1 (NPC1) protein as a novel MP interactor. NPC1 is a transmembrane protein involved in cholesterol transport in animal cells, but also in the infection by several viruses of different families. The identified clone from the melon NPC1 gene spans from exons 25 to 28 and includes two introns. Notably, deletion of the two introns and exon 28 does not impair the interaction capacity of the remaining peptide. The identified CmNPC1 gene maps to chromosome 11. In addition, the melon genome encodes a second copy of NPC1 in chromosome 7 (CmNPC1-C7), highly similar. Functional assays revealed that the interaction domain of CmNPC1-C7 also interacts with CMV MP, suggesting that both genes could have a role in CMV infection. This study represents the first report linking NPC1 to the infection process of a plant virus, expanding our understanding of plant–virus interactions. The Melon Sterol Transporter Niemann-Pick C1 Protein Is a New Interactor of Cucumber mosaic virus Movement Protein 2026-05-20T17:02:04+08:00 LaTeX with hyperref 2026-05-20T12:41:08+02:00 2026-05-20T12:41:08+02:00 Niemann-Pick C1; Cucumber mosaic virus; movement protein; interactor domain; virus infection; melon pdfTeX-1.40.25; modified using OpenPDF 1.4.2 False This is pdfTeX, Version 3.141592653-2.6-1.40.25 (TeX Live 2023) kpathsea version 6.3.5 uuid:d4434b01-2834-4b18-bc07-7072ad7b532f uuid:245275eb-ed82-41fe-9e26-c36dd73b2222 Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 34, 204, 124, 223 Annotation: Subtype: Link Rect: 88, 77, 152, 84 Annotation: Subtype: Link Rect: 34, 64, 119, 72 Annotation: Subtype: Link Rect: 34, 773, 125, 809 Annotation: Subtype: Link Rect: 515, 773, 560, 809 Annotation: Subtype: Link Rect: 97, 663, 108, 677 Annotation: Subtype: Link Rect: 171, 663, 182, 677 Annotation: Subtype: Link Rect: 232, 663, 243, 677 Annotation: Subtype: Link 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