scholarly journals Reconstitution and biochemical characterization of ribonucleoprotein complexes in Type I-E CRISPR–Cas systems

Author(s):  
Yibei Xiao ◽  
Ailong Ke
2008 ◽  
Vol 86 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Zachery R. Belak ◽  
Andrew Ficzycz ◽  
Nick Ovsenek

YY1 (Yin Yang 1) is present in the Xenopus oocyte cytoplasm as a constituent of messenger ribonucleoprotein complexes (mRNPs). Association of YY1 with mRNPs requires direct RNA-binding activity. Previously, we have shown YY1 has a high affinity for U-rich RNA; however, potential interactions with plausible in vivo targets have not been investigated. Here we report a biochemical characterization of the YY1–RNA interaction including an investigation of the stability, potential 5′-methylguanosine affinity, and specificity for target RNAs. The formation of YY1–RNA complexes in vitro was highly resistant to thermal, ionic, and detergent disruption. The endogenous oocyte YY1–mRNA interactions were also found to be highly stable. Specific YY1–RNA interactions were observed with selected mRNA and 5S RNA probes. The affinity of YY1 for these substrates was within an order of magnitude of that for its cognate DNA element. Experiments aimed at determining the potential role of the 7-methylguanosine cap on RNA-binding reveal no significant difference in the affinity of YY1 for capped or uncapped mRNA. Taken together, the results show that the YY1–RNA interaction is highly stable, and that YY1 possesses the ability to interact with structurally divergent RNA substrates. These data are the first to specifically document the interaction between YY1 and potential in vivo targets.


2016 ◽  
Vol 39 (3) ◽  
pp. 397-405 ◽  
Author(s):  
Shaoqing Yang ◽  
Qiaojuan Yan ◽  
Qingdan Bao ◽  
Jingjing Liu ◽  
Zhengqiang Jiang

1996 ◽  
Vol 138 (2-3) ◽  
pp. 147-152 ◽  
Author(s):  
Cheorl-Ho Kim ◽  
Oyekanmi Nashiru ◽  
Jeong Heon Ko

2011 ◽  
Vol 48 (3) ◽  
pp. 260-266 ◽  
Author(s):  
Jinho Kang ◽  
Kyung-Min Park ◽  
Kyoung-Hwa Choi ◽  
Cheon-Seok Park ◽  
Go-Eun Kim ◽  
...  

2018 ◽  
Vol 70 (5-6) ◽  
pp. 1700179 ◽  
Author(s):  
Lingmeng Li ◽  
Fengying Dong ◽  
Lin Lin ◽  
Dannong He ◽  
Jingwen Chen ◽  
...  

2005 ◽  
Vol 70 (3) ◽  
pp. 347-360 ◽  
Author(s):  
Tanja Cirkovic-Velickovic ◽  
Marija Gavrovic-Jankulovic ◽  
Ratko Jankov

The characterization of an allergen is a troublesome and difficult process, as it requires both the precise biochemical characterization of a (glyco)protein molecule and the establishment of its susceptibility to IgE antibodies, as they are the main link to histamine release in some hypersensitivity states (type I allergies). As the characterization of an allergen includes molecular weight determination of the allergenic molecule, its structure determination, physicochemical properties, IgE binding properties of the allergen molecule, and its allergenicity, an overall review of which biochemical and immunochemical methods are used in achieving this goal are presented in this paper. The information on the molecular level on the structures of allergens indicates that allergens are considerably heterogeneous protein structures, and that there is no particular aminoacid sequence which is responsible for the allergenicity. Therefore, information gained from detailed structural, functional and immunochemical studies of these intriguing molecules, which nowadays modulate a variety of pathophysiological conditions, would greatly improve our understanding of the underlying disease mechanisms, and the way to handle them.


2015 ◽  
Vol 108 ◽  
pp. 48-53 ◽  
Author(s):  
Jung-Mi Kang ◽  
Jung-Won Ju ◽  
Jung-Yeon Kim ◽  
Hye-Lim Ju ◽  
Jinyoung Lee ◽  
...  

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