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1.  The long noncoding RNAs NEAT1 and MALAT1 bind active chromatin sites 
Molecular cell  2014;55(5):791-802.
Mechanistic roles for many lncRNAs are poorly understood in part because their direct interactions with genomic loci and proteins are difficult to assess. Using a method to purify endogenous RNAs and their associated factors, we mapped the genomic binding sites for two highly expressed human lncRNAs, NEAT1 and MALAT1. We show that NEAT1 and MALAT1 localize to hundreds of genomic sites in human cells, primarily over active genes. NEAT1 and MALAT1 exhibit colocalization to many of these loci, but display distinct gene body binding patterns at these sites, suggesting independent but complementary functions for these RNAs. We also identified numerous proteins enriched by both lncRNAs, supporting complementary binding and function, in addition to unique associated proteins. Transcriptional inhibition or stimulation alters localization of NEAT1 on active chromatin sites, implying that underlying DNA sequence does not target NEAT1 to chromatin and that localization responds to cues involved in the transcription process.
PMCID: PMC4428586  PMID: 25155612
2.  H3K27 modifications define segmental regulatory domains in the Drosophila bithorax complex 
eLife  2014;3:e02833.
The bithorax complex (BX-C) in Drosophila melanogaster is a cluster of homeotic genes that determine body segment identity. Expression of these genes is governed by cis-regulatory domains, one for each parasegment. Stable repression of these domains depends on Polycomb Group (PcG) functions, which include trimethylation of lysine 27 of histone H3 (H3K27me3). To search for parasegment-specific signatures that reflect PcG function, chromatin from single parasegments was isolated and profiled. The H3K27me3 profiles across the BX-C in successive parasegments showed a ‘stairstep’ pattern that revealed sharp boundaries of the BX-C regulatory domains. Acetylated H3K27 was broadly enriched across active domains, in a pattern complementary to H3K27me3. The CCCTC-binding protein (CTCF) bound the borders between H3K27 modification domains; it was retained even in parasegments where adjacent domains lack H3K27me3. These findings provide a molecular definition of the homeotic domains, and implicate precisely positioned H3K27 modifications as a central determinant of segment identity.
eLife digest
Like other insects, the body of the fruit fly is divided into three main parts—the head, the thorax and the abdomen—and each part, in turn, is made up of several smaller segments. The bithorax complex is a cluster of three genes that together control the identity of the segments that make up the back half of the fruit fly's body. This gene cluster has been studied for several decades and these studies have helped to further our understanding of how genetic information is accessed and used to make an animal’s body plan.
Early on in a fruit fly embryo, stretches of DNA within the bithorax complex regulate where the complex's genes are switched on, and where they are switched off. Proteins called Polycomb group proteins then keep the silenced genes off, in part by adding small chemical marks to other proteins called histones. Most DNA in a cell is wrapped around histones, and the addition of such chemical marks causes the DNA to become more tightly packed. This prevents the bithorax complex genes from being accessed and switched on. It had previously been suggested that each segment might have a unique pattern of chemical marks on the bithorax complex histones, but evidence to support this idea was lacking.
Bowman et al. have now undertaken the technically challenging task of purifying the DNA and its histones from individual segments of fruit fly embryos. This revealed that segments closer to the embryo's head contain larger stretches of bithorax complex DNA covered with histones marked by the Polycomb group proteins. Bowman et al. also found that the coverage of chemical marks on the histones changed dramatically when one segment was compared to its neighboring segments. These sharp boundaries clearly outline which regulatory regions of the DNA are switched on and which are switch off; however the same pattern is not seen for the Polycomb group proteins themselves. Instead, within the bithorax complex, the pattern of these proteins is almost identical in different segments.
The challenge now is to understand how the chemical marks and the Polycomb group proteins work together to restrict access to DNA in such precise patterns. Also—since similar gene clusters control the development of the body plans of mammals—this, in turn, might help us to understand how the Polycomb group proteins perform similar functions in human development and disease.
PMCID: PMC4139060  PMID: 25082344
chromatin; Polycomb; bithorax complex; CTCF; D. melanogaster
3.  A role for central spindle proteins in cilia structure and function 
Cytoskeleton (Hoboken, N.J.)  2011;68(2):112-124.
Cytokinesis and ciliogenesis are fundamental cellular processes that require strict coordination of microtubule organization and directed membrane trafficking. These processes have been intensely studied, but there has been little indication that regulatory machinery might be extensively shared between them. Here, we show that several central spindle/midbody proteins (PRC1, MKLP-1, INCENP, centriolin) also localize in specific patterns at the basal body complex in vertebrate ciliated epithelial cells. Moreover, bioinformatic comparisons of midbody and cilia proteomes reveal a highly significant degree of overlap. Finally, we used temperature-sensitive alleles of PRC1/spd-1 and MKLP-1/zen-4 in C. elegans to assess ciliary functions while bypassing these proteins' early role in cell division. These mutants displayed defects in both cilia function and cilia morphology. Together, these data suggest the conserved re-use of a surprisingly large number of proteins in the cytokinetic apparatus and in cilia.
PMCID: PMC4089984  PMID: 21246755
Ciliogenesis; cytokinesis; PRC1; INCENP; MKLP-1; bioinformatics; cilia midbody
4.  A Census of Human Soluble Protein Complexes 
Cell  2012;150(5):1068-1081.
Cellular processes often depend on stable physical associations between proteins. Despite recent progress, knowledge of the composition of human protein complexes remains limited. To close this gap, we applied an integrative global proteomic profiling approach, based on chromatographic separation of cultured human cell extracts into more than one thousand biochemical fractions which were subsequently analyzed by quantitative tandem mass spectrometry, to systematically identify a network of 13,993 high-confidence physical interactions among 3,006 stably-associated soluble human proteins. Most of the 622 putative protein complexes we report are linked to core biological processes, and encompass both candidate disease genes and unnanotated proteins to inform on mechanism. Strikingly, whereas larger multi-protein assemblies tend to be more extensively annotated and evolutionarily conserved, human protein complexes with 5 or fewer subunits are far more likely to be functionally un-annotated or restricted to vertebrates, suggesting more recent functional innovations.
PMCID: PMC3477804  PMID: 22939629
5.  RIDDLE: reflective diffusion and local extension reveal functional associations for unannotated gene sets via proximity in a gene network 
Genome Biology  2012;13(12):R125.
The growing availability of large-scale functional networks has promoted the development of many successful techniques for predicting functions of genes. Here we extend these network-based principles and techniques to functionally characterize whole sets of genes. We present RIDDLE (Reflective Diffusion and Local Extension), which uses well developed guilt-by-association principles upon a human gene network to identify associations of gene sets. RIDDLE is particularly adept at characterizing sets with no annotations, a major challenge where most traditional set analyses fail. Notably, RIDDLE found microRNA-450a to be strongly implicated in ocular diseases and development. A web application is available at
PMCID: PMC4056375  PMID: 23268829
6.  It’s the machine that matters: Predicting gene function and phenotype from protein networks 
Journal of proteomics  2010;73(11):2277-2289.
Increasing knowledge about the organization of proteins into complexes, systems, and pathways has led to a flowering of theoretical approaches for exploiting this knowledge in order to better learn the functions of proteins and their roles underlying phenotypic traits and diseases. Much of this body of theory has been developed and tested in model organisms, relying on their relative simplicity and genetic and biochemical tractability to accelerate the research. In this review, we discuss several of the major approaches for computationally integrating proteomics and genomics observations into integrated protein networks, then applying guilt-by-association in these networks in order to identify genes underlying traits. Recent trends in this field include a rising appreciation of the modular network organization of proteins underlying traits or mutational phenotypes, and how to exploit such protein modularity using computational approaches related to the internet search algorithm PageRank. Many protein network-based predictions have recently been experimentally confirmed in yeast, worms, plants, and mice, and several successful approaches in model organisms have been directly translated to analyze human disease, with notable recent applications to glioma and breast cancer prognosis.
PMCID: PMC2953423  PMID: 20637909
Data integration; Function prediction; Humans; Model organisms; Phenotype prediction; Protein interaction networks

Results 1-6 (6)