Proximity ligation assays of protein and RNA interactions in the male-specific lethal complex on Drosophila melanogaster polytene chromosomes

Lindehell, Henrik; Kim, Maria; Larsson, Jan
September 2015
Chromosoma;Sep2015, Vol. 124 Issue 3, p385
Academic Journal
In Drosophila, the male-specific lethal (MSL) complex specifically targets the male X chromosome and participates in a twofold increase in expression output leading to functional dosage compensation. The complex includes five proteins and two non-coding RNAs (ncRNAs). A number of additional associated factors have also been identified. However, the components' roles and interactions have not been fully elucidated. The in situ proximity ligation assay (PLA) provides a sensitive means to determine whether proteins and other factors have bound to chromosomes in close proximity to each other, and thus may interact. Thus, we modified, tested, and applied the assay to probe interactions of MSL complex components on polytene chromosomes. We show that in situ PLA can detect and map both protein-protein and protein-ncRNA interactions on polytene chromosomes at high resolution. We further show that all five protein components of the MSL complex are in close proximity to each other, and the ncRNAs roX1 and roX2 bind the complex in close proximity to MLE. Our results also indicate that JIL1, a histone H3 Ser10 kinase enriched on the male X chromosome, interacts with MSL1 and MSL2, but not MSL3 of the MSL complex. In addition, we corroborate proposed interactions of the MSL complex with both CLAMP and TopoII.


Related Articles

  • Inferring Potential microRNA-microRNA Associations Based on Targeting Propensity and Connectivity in the Context of Protein Interaction Network. Sun, Jie; Zhou, Meng; Yang, Haixiu; Deng, Jiaen; Wang, Letian; Wang, Qianghu // PLoS ONE;Jul2013, Vol. 8 Issue 7, p1 

    MicroRNAs (miRNAs) are a group of small non-coding RNAs that play important regulatory roles at the post-transcriptional level. Although several computational methods have been developed to compare miRNAs, it is still a challenging and a badly needed task with the availability of various...

  • Long Noncoding RNA MEG3 Interacts with p53 Protein and Regulates Partial p53 Target Genes in Hepatoma Cells. Zhu, Juanjuan; Liu, Shanshan; Ye, Fuqiang; Shen, Yuan; Tie, Yi; Zhu, Jie; Wei, Lixin; Jin, Yinghua; Fu, Hanjiang; Wu, Yongge; Zheng, Xiaofei // PLoS ONE;10/7/2015, Vol. 10 Issue 10, p1 

    Maternally Expressed Gene 3 (MEG3) encodes a lncRNA which is suggested to function as a tumor suppressor. Previous studies suggested that MEG3 functioned through activation of p53, however, the functional properties of MEG3 remain obscure and their relevance to human diseases is under continuous...

  • Role of GW182 proteins and PABPC1 in the miRNA pathway: a sense of déjà vu. Tritschler, Felix; Huntzinger, Eric; Izaurralde, Elisa // Nature Reviews Molecular Cell Biology;May2010, Vol. 11 Issue 5, p379 

    GW182 proteins have emerged as key components of microRNA (miRNA) silencing complexes in animals. Although the precise molecular function of GW182 proteins is not fully understood, new findings indicate that they act as poly(A)-binding protein (PABP)-interacting proteins (PAIPs) that promote...

  • Integrative network analysis reveals active microRNAs and their functions in gastric cancer. Chien-Wei Tseng; Chen-Ching Lin; Chiung-Nien Chen; Hsuan-Cheng Huang; Hsueh-Fen Juan // BMC Systems Biology;2011, Vol. 5 Issue 1, p99 

    Background: MicroRNAs (miRNAs) are a class of endogenous, small and highly conserved noncoding RNAs that control gene expression either by degradation of target mRNAs or by inhibition of protein translation. They play important roles in cancer progression. A single miRNA can provoke a chain...

  • Bacterial Small Regulatory RNAs and Hfq Protein. Murina, V.; Nikulin, A. // Biochemistry (00062979);Dec2015, Vol. 80 Issue 13, p1647 

    Small regulatory RNA (sRNA) is a unique noncoding RNA involved in regulation of gene expression in both eukaryotic and bacterial cells. This short review discusses examples of positive and negative translation regulation by sRNAs in bacteria and participation of Hfq in these processes. The...

  • MicroRNAs in control of cardiac hypertrophy. Da Costa Martins, Paula A.; De Windt, Leon J. // Cardiovascular Research;Mar2012, Vol. 93 Issue 4, p563 

    MicroRNAs refer to a subfamily of small non-coding RNA species that are designed to influence gene expression in nearly all cell types studied to date. Studies from the past decade have demonstrated that microRNAs are atypically expressed in the cardiovascular system under specific pathological...

  • MicroRNA-451 functions as a tumor suppressor in human non-small cell lung cancer by targeting ras-related protein 14 (RAB14). Wang, R; Wang, Z-X; Yang, J-S; Pan, X; De, W; Chen, L-B // Oncogene;6/9/2011, Vol. 30 Issue 23, p2644 

    Accumulating evidence suggests that microRNAs (miRNAs) are important gene regulators, which can have critical roles in diverse biological processes including tumorigenesis. In this study, we analyzed the miRNA expression profiles in non-small cell lung carcinoma (NSCLC) by use of a miRNA...

  • microRNA Replacement Therapy for Cancer. Henry, Jon; Azevedo-Pouly, Ana; Schmittgen, Thomas // Pharmaceutical Research;Dec2011, Vol. 28 Issue 12, p3030 

    microRNA are small noncoding RNAs that translationally repress their target messenger RNAs. Many microRNAs are expressed at reduced levels in tumors. microRNAs with reduced expression in cancer often regulate oncogenes, resulting in enhanced tumor growth. One therapeutic option is to restore...

  • Target Gene and Function Prediction of Differentially Expressed MicroRNAs in Lactating Mammary Glands of Dairy Goats. Fei Dong; Zhi-Bin Ji; Cun-Xian Chen; Gui-Zhi Wang; Jian-Min Wang // International Journal of Genomics;2013, p1 

    MicroRNAs are small noncoding RNAs that can regulate gene expression, and they can be involved in the regulation of mammary gland development. The differential expression of miRNAs during mammary gland development is expected to provide insight into their roles in regulating the homeostasis of...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics