Difference between revisions of "Part:BBa K1150001"
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| + | VP16 | ||
Virus Protein 16 (VP16) is a transcription factor encoded by the UL48 gene of Herpes simplex virus-1 (HSV-1). | Virus Protein 16 (VP16) is a transcription factor encoded by the UL48 gene of Herpes simplex virus-1 (HSV-1). | ||
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'''Fig. 1:''' Domain structure of VP16. The different functional domains of VP16 are shown. Taken from Hirai et al., 2010. | '''Fig. 1:''' Domain structure of VP16. The different functional domains of VP16 are shown. Taken from Hirai et al., 2010. | ||
| + | ===Usage and Biology=== | ||
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The TAD of VP16 is one of the most efficient TADs. It is widely fused to host transcription factors in order to increase their activity or, next to this, to other transcription factors to study the mechanism of gene regulation [Hirai et al., 2010]. | The TAD of VP16 is one of the most efficient TADs. It is widely fused to host transcription factors in order to increase their activity or, next to this, to other transcription factors to study the mechanism of gene regulation [Hirai et al., 2010]. | ||
In our case the TAD of VP16 was cloned behind a double mutated Cas9 gene (which encodes for a non-cutting Cas9 protein with specific DNA binding properties) to activate our genes of interest. For more information see [[BBa_K1150019]] and [[BBa_K1150020]]. | In our case the TAD of VP16 was cloned behind a double mutated Cas9 gene (which encodes for a non-cutting Cas9 protein with specific DNA binding properties) to activate our genes of interest. For more information see [[BBa_K1150019]] and [[BBa_K1150020]]. | ||
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| + | ==Biology== | ||
| + | VP16 is originally a part of the virus particle from HSV and is released into an animal cell while infection. VP16 first binds via its core domain to the host nuclear protein which then recruits another host nuclear protein, Oct-1, to form a functional protein complex. This complex then binds to | ||
| + | its target DNA sequence in the promotor region of immediate-early genes where VP16 activates these | ||
| + | genes through interactions between its TAD and many different | ||
| + | transcription factors and other proteins, such as histone acetyltransferases; see Fig.2. Through these multiple interactions the transactivation domain of VP16 facilitates the assembly of the pre-initiation complex [Walker et al., 1993; Hirai et al., 2010]. Also other TAD containing proteins share this magnitude of binding partners but VP16 is one of the most effective once [ Hirai et al., 2010]. | ||
Revision as of 12:40, 26 September 2013
VP16
VP16
Virus Protein 16 (VP16) is a transcription factor encoded by the UL48 gene of Herpes simplex virus-1 (HSV-1).
Through complex formation with cellular host factors VP16 can bind to a common regulatory element in the upstream promotor region of immediate-early genes [Weir, 2001]. Through the transactivating function of VP-16 these genes can then be expressed.
VP16 consists of 490 amino acids with two very important functional domains: a core domain in its central region which is necessary for the indirect DNA binding and a carboxy-terminal transactivation domain (Greaves and O’Hare, 1989; Triezenberg et al., 1988); see Fig.1. The transactivation domain (TAD) of VP16 can be fused to a DNA-binding domain (DBD) of another protein in order to gain expression of a desired target gene [Hirai et al., 2010].
Fig. 1: Domain structure of VP16. The different functional domains of VP16 are shown. Taken from Hirai et al., 2010.
Usage and Biology
Sequence and Features
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