Melee:saga expt

20.109 Module 3: Expression Engineering

Primer Design

Forward Primer:
5' aaaagtcttcagttaactcaggttcgtattctacattagATGTCGAAAGCTACATATAA 3'
Reverse Primer:
5' cttcgaaaggaatagtagcggaaaagcttcttctacgcaTTAGTTTTGCTGGCCGCATC 3'

SAGA subunits

Ada subunits	size,chromosome,null p-type	notes
Ada1 (aka HFI1, SUP110, SRM12, GAN1)	1.467 kb/489 aa, Chr. XVI, viable	Adaptor protein required for structural integrity of the SAGA complex, a histone acetyltransferase-coactivator complex that is involved in global regulation of gene expression through acetylation and transcription functions exhibits growth defect on a non-fermentable (respiratory) carbon source
Ada2 (aka SWI8)	1.305 kb/434aa, Chr. IV, viable	Transcription coactivator, component of the ADA and SAGA transcriptional adaptor/HAT (histone acetyltransferase) complexes decreased drug resistance (rapamycin) decreased drug resistance (wortmannin)
Ada3(aka NGG1, SWI7)	2.109 kb/702aa, Chr. IV, viable	Transcriptional regulator involved in glucose repression of Gal4p-regulated genes; component of transcriptional adaptor and histone acetyltransferase complexes, the ADA complex, the SAGA complex, and the SLIK complex decreased drug resistance (rapamycin) decreased drug resistance (wortmannin)
Gcn5 (aka ADA4, SWI9)	1.32 kb/439aa, Chr. VII, viable	Histone acetyltransferase, acetylates N-terminal lysines on histones H2B and H3; catalytic subunit of the ADA and SAGA histone acetyltransferase complexes; founding member of the Gcn5p-related N-acetyltransferase superfamily
Ada5 (aka SPT20)	1.815 kb/604aa, Chr. XV, viable	Subunit of the SAGA transcriptional regulatory complex, involved in maintaining the integrity of the complex decreased drug resistance (rapamycin) decreased drug resistance (wortmannin)

Spt subunits	size, chromosome, null p-type	notes
Spt3	1.014 kb/337aa, Chr. IV, viable	Subunit of the SAGA and SAGA-like transcriptional regulatory complexes, interacts with Spt15p to activate transcription of some RNA polymerase II-dependent genes, also functions to inhibit transcription at some promoters increased drug resistance (rapamycin) decreased drug resistance (wortmannin)
Spt7(aka GIT2)	3.999 kb/1332aa, Chr. II, viable	Subunit of the SAGA transcriptional regulatory complex, involved in proper assembly of the complex; also present as a C-terminally truncated form in the SLIK/SALSA transcriptional regulatory complex increased drug resistance (rapamycin) decreased drug resistance (wortmannin)
Spt8	1.809 kb/602aa, Chr. XII, viable	Subunit of the SAGA transcriptional regulatory complex but not present in SAGA-like complex SLIK/SALSA, required for SAGA-mediated inhibition at some promoters Exhibits sensitivity at 5 generations when grown in synthetic complete - trp medium. normal chitin levels increased drug resistance (rapamycin) decreased drug resistance (wortmannin)
Spt20 (aka Ada5)	1.815 kb/604aa, Chr. XV, viable	Subunit of the SAGA transcriptional regulatory complex, involved in maintaining the integrity of the complex decreased drug resistance (rapamycin) decreased drug resistance (wortmannin)

TAF subunits	size, chromosome, null p-type	notes
TAF5 (aka TAF90)	2.397 kb/798aa, Chr. II, inviable	Subunit (90 kDa) of TFIID and SAGA complexes, involved in RNA polymerase II transcription initiation and in chromatin modification
TAF6 (aka TAF60)	1.551 kb/516aa, Chr. VII, inviable	Subunit (60 kDa) of TFIID and SAGA complexes, involved in transcription initiation of RNA polymerase II and in chromatin modification, similar to histone H4
TAF9 (aka TAF17)	0.474 kb/157aa, Chr. XIII, inviable	Subunit (17 kDa) of TFIID and SAGA complexes, involved in RNA polymerase II transcription initiation and in chromatin modification, similar to histone H3
TAF10 (aka TAF23, TAF25)	0.621 kb/206aa, Chr. IV, inviable	Subunit (145 kDa) of TFIID and SAGA complexes, involved in RNA polymerase II transcription initiation and in chromatin modification
TAF12(aka TAF61, TAF68)	1.620 kb/539aa, Chr. IV, inviable	Subunit (61/68 kDa) of TFIID and SAGA complexes, involved in RNA polymerase II transcription initiation and in chromatin modification, similar to histone H2A

Tra1 subunit	size, chromosome, null p-type	notes
Tra1	11.235 kb/3744aa, Chr. VIII, inviable	Subunit of SAGA and NuA4 histone acetyltransferase complexes; interacts with acidic activators (e.g., Gal4p) which leads to transcription activation; similar to human TRRAP, which is a cofactor for c-Myc mediated oncogenic transformation

other subunits	size, chromosome, null p-type	notes
Sgf73	1.974 kb/657aa, Chr. VII , viable	73 kDa subunit of SAGA histone acetyltransferase complex; involved in formation of the preinitiation complex assembly at promoters
Sgf29	0.779 kb/259aa, Chr. III, viable	SaGa associated Factor 29kDa; Probable 29kKDa Subunit of SAGA histone acetyltransferase complex Exhibits sensitivity at 15 generations when grown in medium of pH 8.
Sgf11	0.3 kb/99aa, Chr.XVI, viable	Integral subunit of SAGA histone acetyltransferase complex, regulates transcription of a subset of SAGA-regulated genes, required for the Ubp8p association with SAGA and for H2B deubiquitylation
Ubp8	1.416 kb/471aa, Chr. XIII, viable	Ubiquitin-specific protease that is a component of the SAGA acetylation complex; required for SAGA-mediated deubiquitination of histone H2B
Sus1	gene with intron, Chr. II, viable	Protein involved in mRNA export coupled transcription activation; component of the SAGA histone acetylase complex