Chromosome-Nuclear Envelope Interaction

Chromosome-Nuclear Envelope Interaction proteins (Lieb project, Strome subgroup)

General Description

The focus of our analysis will be elements that specify nucleosome positioning and occupancy, control domains of gene expression, induce repression of the X chromosome, guide mitotic segregation and genome duplication, govern homolog pairing and recombination during meiosis, and organize chromosome positioning within the nucleus. 126 strategically selected targets include key histone modifications, histone variants, RNA polymerase II isoforms, dosage-compensation proteins, centromere components, homolog-pairing facilitators, recombination markers, and nuclear-envelope constituents.
We will integrate information generated with existing knowledge on the biology of the targets, perform ChIP-chip analysis on mutant and RNAi extracts lacking selected target proteins, use extrachromosomal arrays to assess the ability of candidate identified sequence motifs to recruit targets in vivo, identify tissue-specific patterns of selected targets, and create integrated, quantitative models of transcription and whole-chromosome functions.

Protocols

  1. Growth and isolation: Worm embryo growth and harvest:JL:5, Worm embryo extraction:JL:2
  2. Sample preparation: Worm LM-PCR Amplification for ChIP-chip:JL:KI3, Worm chromatin immunoprecipitation:JL:KI3, ChIP-chip scanning nimblegen:JL:2, ChIP-chip label hyb nimblegen:JL:2
  3. Other Protocols: ChIP-chip normalization standard MA2C:JL:2

Experimental Reagents

    Growth Conditions:
  1. Antibodies: SDQ4094_NPP13, SDQ4051_LEM2, SDQ3891_LEM2
  2. Arrays: 080922_MODENCODE_CE_CHIP_HX1

Sample Details

  1. Animals/Lines: Mixed embryos 20dC

Release Date: 2010-01-31