#235
Vesicular Trafficking in Root Tips

Stephanie Robert (P.I.: Natasha Raikhel and Zhenbiao Yang)

Jae-Ung Hwang and Shundai Li

It is known that there are genetic links between auxin transport, root growth and the endomembrane system. The screen was to examine the root growth when the seedlings are treated with chemicals and then observe PIN2-GFP localization under short time chemical treatment. SCREEN DETAILS: For the primary screen a total of 42 000 diverse chemicals were screened. The libraries were: (1. Sigma TimTec Myria Screen (MS) (10,000), 2. Chembridge Nova Core Library (NC) (10,000), 3. Selected chemicals from Microsource Spectrum (SP) (2 000), 4. Chembridge Diverset (ND) (20 000) ). The SP,MS, NC and ND libraries were screened at a concentration of 50-100 uM in 400 ul of medium using standard 24-well microtiter plates. Seeds were germinated on media with chemicals. Chemicals that inhibited root growth were selected after 4 to 5 days. For the secondary screen, seedlings expressing PIN2-GFP and markers for different endomembrane compartments were germinated and grown for 4-5 days and treated for 2 hrs with chemicals at concentrations below those of the primary screen. Overall, only compounds that altered marker localization and inhibited root growth were selected. SUMMARY STATISTICS: Of the 42,000 chemicals screened, a total of 1721 (4.1%). In the secondary screen, 11 of the 1721 chemicals (0.03% of the 42,000) resulted in altered PIN2-GFP localization/expression. 4 of these 11 (0.01% of the 42,000) were confirmed to alter the localization/expression of PIN2-GFP or different endomembrane markers.

NSF: 2010-0520325

Visiable to everybody

  • Forward Screenyes
  • Target Pathwayvesicular trafficking, auxin transport, cell elongation
  • Screening Material
    • OrganismArabidopsis thaliana
    • EcotypeColumbia (Col-0)
    • GenotypePIN2::PIN2-GFP
    • Tissue and/or Cell Type(s)root tip, epidermal and cortical cells
    • Sample Age4-5 days
    • Growth Conditionssterilized seeds were grown on MS medium in light conditions (light 16hrs 22.5C, night 8hrs 21C, on MS medium + 1% sucrose)and treated with chemicals from 30 min to 2h

In total, 4 compounds from 1 library are annotated. Out of these, 2 have a score of 1 or higher, and 0 are annotated as inactive.

Download Download Compound IDs

Screen-level Files

Original Pin2 Publication

#236
Muller A, Guan C, Galweiler L, Tanzler P, Huijser P, Marchant A, Parry G, Bennett M, Wisman E, Palme K. (1998), AtPIN2 defines a locus of Arabidopsis for root gravitropism control., appeared in EMBO, 17, 6903-6911., pp 17, 6903-6911..

Screen Strategy

#15325
Download

Compound Files

ChemBridge Diverset 20K 6534658
LibraryChemBridge Diverset 20K
ID6534658
Name
FormulaC19H21ClFN3O
Molecular Weight 361.841
Use in structure search

Assay 1: 3

#8746
Assay 1: visual phenotype
  • Description None
  • Score 3: active in all previous and follow-up screens
  • Concentration 50-100 uM (25 ug/ml) in 0.5% DMSO

6534658-PIN2-GFP

'''PIN2''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of PIN2-GFP, agglomerations

#15293
Main image
Reference
Download

6534658-ARA7-GFP

'''ARA7''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of ARA7-GFP

#15294
Main image
Reference
Download

6534658-dTIP-GFP

'''Delta-TIP''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of Delta-TIP-GFP, cytosolic

#15295
Main image
Reference
Download

6534658-Photo1-GFP

'''Photo1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of Phototropin1-GFP

#15296
Main image
Reference
Download

6534658-SNX1-GFP

'''SNX1''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of SNX1-GFP, weak signal, cytosolic

#15297
Main image
Reference
Download

6534658-PIN1-GFP

'''PIN1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of PIN1-GFP

#15298
Main image
Reference
Download
ChemBridge Diverset 20K 6077129
LibraryChemBridge Diverset 20K
ID6077129
Name
FormulaC15H14FNO2S
Molecular Weight 291.341
Use in structure search

Standard Compound Annotation
Assay 1: 3

#117186
Assay 1: visual phenotype
  • Score 3: active in all previous and follow-up screens
  • Concentration 50-100 uM (25 ug/ml) in 0.5% DMSO

6077129-PIN2-GFP

'''PIN2''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of PIN2-GFP, bigger endosomes

#15303
Main image
Reference
Download

6077129-ARA7-GFP

'''ARA7''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of ARA7-GFP but the signal is weaker

#15304
Main image
Reference
Download

6077129-dTIP-GFP

'''Delta-TIP''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of Delta-TIP-GFP

#15305
Main image
Reference
Download

6077129-Photo1-GFP

'''Photo1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of Phototropin1-GFP

#15306
Main image
Reference
Download

6077129-SNX1-GFP

'''SNX1''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of SNX1-GFP, weaker signal

#15307
Main image
Reference
Download

6077129-PIN1-GFP

'''PIN1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of PIN1-GFP

#15308
Main image
Reference
Download
ChemBridge Diverset 20K 6538980
LibraryChemBridge Diverset 20K
ID6538980
Name
FormulaC17H15ClFNO3
Molecular Weight 335.757
Use in structure search

6538980-PIN2-GFP

'''PIN2''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of PIN2-GFP, agglomerations

#15309
Main image
Reference
Download

6538980-Photo1-GFP

'''Photo1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of Phototropin1-GFP

#15311
Main image
Reference
Download
ChemBridge Diverset 20K 5711337
LibraryChemBridge Diverset 20K
ID5711337
Name
FormulaC19H26N2O2
Molecular Weight 314.422
Use in structure search

5711337-PIN2-GFP

'''PIN2''', 25-50 um chemical (left) compared to DMSO control (right). Results: mislocalization of PIN2-GFP

#15312
Main image
Reference
Download

5711337-Photo1-GFP

'''Photo1''', 25-50 um chemical (left) compared to DMSO control (right). Results: normal localization of Phototropin1-GFP

#15313
Main image
Reference
Download

Brought to you by UCR::IIGB::CEPCEB