TABLE 2.

Discrimination values for ribonucleotide analogs by POLRMT^a

Nucleotide analog	${\mathit{D}*}_{\mathbf{\text{analog}}}$ (mean ± SD)	${{\mathit{D}}^{\mathbf{\text{cal}}}}_{\mathbf{\text{analog}}}$ (mean ± SD)	Misincorporation frequency
ATP analogs
3′-dATP	1	21 ± 0.4	(4.8 ± 0.09) × 10−2
2′-dATP	375 ± 57	7,875 ± 1,197	(1.3 ± 0.2) × 10−4
2′-F-ATP	240 ± 17	5,040 ± 357	(2.0 ± 0.1) × 10−4
7-deaza-ATP		2.95 ± 0.07	(3.4 ± 0.08) × 10−1
2′-C-Me-ATP	123 ± 8	2,583 ± 168	(3.9 ± 0.3) × 10−4
2′-C-ethynyl-7-deaza-ATP	107 ± 4	2,247 ± 84	(4.5 ± 0.2) × 10−4
ara-ATP	226 ± 16	4,746 ± 336	(2.1 ± 0.1) × 10−4
GTP analogs
3′-dGTP	1	34.5 ± 0.8	(2.9 ± 0.07) × 10−2
2′-dGTP	82.2 ± 8.3	2,836 ± 286	(3.5 ± 0.4) × 10−4
2′-F-GTP	44.7 ± 4.7	1,542 ± 162	(6.5 ± 0.7) × 10−4
2′-C-Me-GTP	83.0 ± 8.9	2,864 ± 307	(3.5 ± 0.4) × 10−4
2′-F-2′-C-Me-GTP	∼7,618 ± 3,042	∼262,821 ± 104,949	(∼3.8 ± 1.5) × 10−6
2′-NH2-GTP	2.0 ± 0.4	69 ± 13.8	(1.4 ± 0.3) × 10−2
2′-azido-GTP	115 ± 13	3,968 ± 449	(2.5 ± 0.3) × 10−4
ara-GTP	18.8 ± 0.1	649 ± 3	(1.5 ± 0.007) × 10−3
4′-azido-GTP	0.6 ± 0.1	20.7 ± 3.45	(4.8 ± 0.8) × 10−2
CTP analogs
3′-dCTP	1	59.7 ± 1.6	(1.7 ± 0.04) × 10−2
2′-dCTP	77.2 ± 3.5	4,609 ± 209	(2.2 ± 0.1) × 10−4
2′-F-CTP	22.7 ± 0.2	1,355 ± 12	(7.4 ± 0.07) × 10−4
2′-NH2-CTP	1.2 ± 0.1	71.6 ± 6	(1.4 ± 0.1) × 10−2
2′-azido-CTP	32.8 ± 1.8	1,958 ± 107	(5.1 ± 0.3) × 10−4
Ara-CTP	241 ± 8	14,388 ± 478	(7.0 ± 0.2) × 10−5
2′-F-2′-C-Me-CTP	∼3,569 ± 504	∼213,069 ± 30,089	(∼4.7 ± 0.7) × 10−6
2′-C-Me-CTP	116 ± 7	6,925 ± 418	(1.4 ± 0.09) × 10−4
4′-azido-CTP	44.9 ± 4.1	2,681 ± 245	(3.7 ± 0.3) × 10−4
UTP analogs
3′-dUTP	1	94.7 ± 8	(1.1 ± 0.09) × 10−2
2′-dUTP	10.6 ± 4.7	1,004 ± 445	(1.0 ± 0.4) × 10−3
2′-dTTP	35.2 ± 7.8	3,333 ± 739	(3.0 ± 0.7) × 10−4
2′-F-UTP	7.4 ± 1.6	701 ± 152	(1.4 ± 0.3) × 10−3
2′-F-2′-C-Me-UTP	No incorporation	NA	NA
2′-C-Me-UTP	714 ± 80	67,616 ± 7,576	(1.5 ± 0.2) × 10−5
2′-C-ethynyl-UTP	697 ± 50	66,006 ± 4,735	(1.5 ± 0.1) × 10−5
ara-UTP	∼1,173 ± 135	∼111,083 ± 12,785	(∼9.0 ± 1) × 10−6
4′-azido-UTP	3.5 ± 0.8	331 ± 76	(3.0 ± 0.7) × 10−3

^a${D*}_{\text{analog}}=K_{1/2,\text{analog\hspace{0.17em}NTP}}/K_{1/2,3\prime \text{-}\text{dNTP}};{D^{cal}}_{\text{analog}}={D*}_{\text{analog}}\times D_{3\prime \text{-dNTP}}$, where the values of D_3′-dNTP are from Table 1. Misincorporation efficiency is equal to $1/{D^{cal}}_{\text{analog}}$. The data are from 2 or 3 independent experiments.