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Anti-FLAG-affinity purification of Umbrea associated factors

ABSTRACT: Anti-FLAG-affinity purification of Umbrea associated factors from nuclear extracts of Drosophila Schneider cells expressing FLAG-HA-Umbrea. Anti-FLAG -affinity purifications from Schneider cell nuclear extracts not expressing any FLAG-tagged protein served as a control SDS-PAGE separting both samples, 9 slices per sample, reduction and alkylation, digestion with trypsin. LC-MS on a Ultimate3000-LTQ Orbitrap XL Raw data were analysed using the MaxQuant 1.2.2.5 software package. Identified proteins were considered as interaction partners if their MaxQuant intensities displayed a greater than 10 fold enrichment compared to control anti-FLAG purifications from L2-4 nuclear extracts not expressing any FLAG-tagged protein

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

SUBMITTER: Andreas Thomae

PROVIDER: PXD000163 | Pride | 2013-10-03

REPOSITORIES: Pride

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Stepwise evolution of essential centromere function in a Drosophila neogene.

Ross Benjamin D BD Rosin Leah L Thomae Andreas W AW Hiatt Mary Alice MA Vermaak Danielle D de la Cruz Aida Flor A AF Imhof Axel A Mellone Barbara G BG Malik Harmit S HS

Science (New York, N.Y.) 20130601 6137

Evolutionarily young genes that serve essential functions represent a paradox; they must perform a function that either was not required until after their birth or was redundant with another gene. How young genes rapidly acquire essential function is largely unknown. We traced the evolutionary steps by which the Drosophila gene Umbrea acquired an essential role in chromosome segregation in D. melanogaster since the gene's origin less than 15 million years ago. Umbrea neofunctionalization occurre ...[more]

PMID: 23744945

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